Biochemical Engineering Journal最新文献_第8页

Stepwise enhancement of selectivity in hydrophobic charge-induction chromatography through grafted and dual-ligand systems 通过接枝和双配体体系逐步增强疏水电荷感应色谱的选择性

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-11-04 DOI: 10.1016/j.bej.2025.109991

Wei Shi , Hong-Wen Zhou , Sha-Sha Deng , Yu-Qian Dai , Si-Qi Zhang , Kai-Bin Li , Xiao-Bin Zhang , De-Man Han

The separation of protein-based therapeutics from complex feedstocks remains a major challenge, especially when high selectivity among proteins differing in size and conformation is required. Hydrophobic Charge-Induction Chromatography (HCIC) combines hydrophobic interactions with pH-dependent charge induction, enabling mild elution and activity retention. Here, we present a stepwise strategy to enhance selectivity in HCIC: (i) introducing a grafted polymer layer (grafted single-ligand system) and (ii) combining grafting with a second ligand (grafted dual-ligand system) using ligands 2-(aminomethyl)pyridine and 4-(1H-imidazol-1-yl)aniline. Comparative studies of ungrafted single-ligand (4FF-S), grafted single-ligand (4FF-G-S), and grafted dual-ligand (4FF-G-D) resins were performed using bovine immunoglobulin G (bIgG) and bovine serum albumin (BSA) as model proteins. Characterization confirmed structural modifications, yielding average pore sizes of 13.1, 20.5, and 19.8 nm for 4FF-S120, 4FF-G130-S250, and 4FF-G130-D250 + 140, respectively. Under optimal conditions (pH 7.0, no salt), selectivity factors (α = Q_bIgG/Q_BSA) increased from 1.5 to 3.7 and 6.8, corresponding to 146 % and 353 % improvements relative to the baseline. Adsorption kinetics, breakthrough and serum separation experiments corroborated that grafting enhances preferential bIgG binding, while the dual-ligand design further amplifies selectivity through synergistic multi-site interactions. Correspondingly, the bIgG recovery improved from 69.5 % to 85.5 % and 92.7 %. These findings demonstrate that grafted and dual-ligand systems are effective and selective under dynamic conditions, offering a practical and scalable route for precise protein purification in biopharmaceutical processes.

从复杂的原料中分离基于蛋白质的治疗方法仍然是一个主要的挑战，特别是当需要对不同大小和构象的蛋白质进行高选择性时。疏水电荷诱导色谱（HCIC）结合了疏水相互作用和ph依赖的电荷诱导，实现了温和的洗脱和活性保留。在这里，我们提出了一种逐步提高HCIC选择性的策略：(i)引入接枝聚合物层（接枝单配体体系）；（ii）使用配体2-（氨基甲基）吡啶和4-（1h -咪唑-1-基）苯胺与第二配体（接枝双配体体系）结合接枝。以牛免疫球蛋白G （bIgG）和牛血清白蛋白（BSA）为模型蛋白，对未接枝的单配体（4FF-S）、接枝的单配体（4FF-G-S）和接枝的双配体（4FF-G-D）树脂进行了比较研究。表征证实了结构的改变，4FF-S120、4FF-G130-S250和4FF-G130-D250的平均孔径分别为13.1、20.5和19.8 nm + 140。在最佳条件下（pH 7.0，无盐），选择性因子（α = QbIgG/QBSA）从1.5增加到3.7和6.8，相对于基线分别提高了146 %和353 %。吸附动力学、突破和血清分离实验证实，接枝增强了bIgG的优先结合，而双配体设计通过协同多位点相互作用进一步增强了选择性。相应的，bIgG回收率从69.5 %提高到85.5 %和92.7 %。这些发现表明，在动态条件下，接枝和双配体系统是有效和选择性的，为生物制药过程中的精确蛋白质纯化提供了一条实用和可扩展的途径。

{"title":"Stepwise enhancement of selectivity in hydrophobic charge-induction chromatography through grafted and dual-ligand systems","authors":"Wei Shi , Hong-Wen Zhou , Sha-Sha Deng , Yu-Qian Dai , Si-Qi Zhang , Kai-Bin Li , Xiao-Bin Zhang , De-Man Han","doi":"10.1016/j.bej.2025.109991","DOIUrl":"10.1016/j.bej.2025.109991","url":null,"abstract":"<div><div>The separation of protein-based therapeutics from complex feedstocks remains a major challenge, especially when high selectivity among proteins differing in size and conformation is required. Hydrophobic Charge-Induction Chromatography (HCIC) combines hydrophobic interactions with pH-dependent charge induction, enabling mild elution and activity retention. Here, we present a stepwise strategy to enhance selectivity in HCIC: (i) introducing a grafted polymer layer (grafted single-ligand system) and (ii) combining grafting with a second ligand (grafted dual-ligand system) using ligands 2-(aminomethyl)pyridine and 4-(1H-imidazol-1-yl)aniline. Comparative studies of ungrafted single-ligand (4FF-S), grafted single-ligand (4FF-G-S), and grafted dual-ligand (4FF-G-D) resins were performed using bovine immunoglobulin G (bIgG) and bovine serum albumin (BSA) as model proteins. Characterization confirmed structural modifications, yielding average pore sizes of 13.1, 20.5, and 19.8 nm for 4FF-S120, 4FF-G130-S250, and 4FF-G130-D250 + 140, respectively. Under optimal conditions (pH 7.0, no salt), selectivity factors (α = <em>Q</em><sub>bIgG</sub>/<em>Q</em><sub>BSA</sub>) increased from 1.5 to 3.7 and 6.8, corresponding to 146 % and 353 % improvements relative to the baseline. Adsorption kinetics, breakthrough and serum separation experiments corroborated that grafting enhances preferential bIgG binding, while the dual-ligand design further amplifies selectivity through synergistic multi-site interactions. Correspondingly, the bIgG recovery improved from 69.5 % to 85.5 % and 92.7 %. These findings demonstrate that grafted and dual-ligand systems are effective and selective under dynamic conditions, offering a practical and scalable route for precise protein purification in biopharmaceutical processes.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109991"},"PeriodicalIF":3.7,"publicationDate":"2025-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Study on structural analysis, decomposition reaction kinetics and extraction separation of green fluorescent substances in caffeine synthesis 咖啡因合成中绿色荧光物质的结构分析、分解反应动力学及萃取分离研究

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-11-03 DOI: 10.1016/j.bej.2025.109990

Binbin Zhao , Jincai Yue , Bo Zhou , Lingqi Kong

Caffeine is a widely used psychoactive substance. The green fluorescent substance (GFS) generated during its chemical synthesis seriously affects the appearance of products, leading to off-specification batches and increased wastewater. This study purified GFS to 95.87 % purity via preparative high-performance liquid chromatography (HPLC), identifying it as 1,3-dimethyl-7-(methylamino)-2,4-dioxo-1,2,3,4-tetrahydropteridine-6-carboxylic acid (C₁₀H₁₁N₅O₄). Quantitative analysis showed GFS forms during violuric acid hydrogenation, with partial decarboxylation in the closed-loop stage but fluctuating residues that compromise quality. Based on these findings, experimental studies were conducted on the decomposition behavior of GFS under alkaline conditions and its extraction removal characteristics. Chloroform, a polar solvent, has high extraction rate for GFS. Extraction rate increased with decreasing pH and temperature, reaching 74.92 % at pH 2 and 308.15 K when the same mass of extractant as the GFS aqueous solution was added. Decomposition reaction studies demonstrated that the decomposition rate

x_{GFS}

increased significantly with higher NaOH concentration and reaction temperature; under conditions of 8 mol·L⁻¹ NaOH, 368.15 K, and 50 min,

x_{GFS}

reached 100 %. Regression analysis yielded the following kinetic equation for the decomposition reaction under alkaline conditions:

r = - \exp (\frac{55.32 - 4.72 m}{RT} + 10.82 - 1.47 m) C_{A}

. This provides critical data for a "solvent extraction + alkaline decomposition" process to improve caffeine production and quality, and reduce wastewater discharge.

咖啡因是一种广泛使用的精神活性物质。其化学合成过程中产生的绿色荧光物质（GFS）严重影响了产品的外观，导致批次不合格，废水增加。本研究通过制备型高效液相色谱（HPLC）纯化GFS纯度为95.87 %，鉴定其为1,3-二甲基-7-（甲氨基）-2,4-二氧基-1,2,3,4-四氢叔丁基-6-羧酸（C₁₀H₁₁N₅O₄）。定量分析表明，在紫尿酸加氢过程中，GFS形成，在闭环阶段部分脱羧，但波动残基影响质量。在此基础上，对GFS在碱性条件下的分解行为及其萃取去除特性进行了实验研究。氯仿是一种极性溶剂，对GFS有较高的提取率。萃取率随pH和温度的降低而增加，在pH为2时达到74.92 %，在加入与GFS水溶液质量相同的萃取剂时达到308.15 K。分解反应研究表明，随着NaOH浓度和反应温度的升高，分解速率xGFS显著增加；在8mol·L⁻¹NaOH， 368.15 K， 50min的条件下，xGFS达到100%。通过回归分析得到碱条件下分解反应的动力学方程：r= - exp(55.32−4.72mRT+10.82−1.47m)CA。这为“溶剂萃取+碱性分解”工艺提供了关键数据，以提高咖啡因的产量和质量，并减少废水排放。

{"title":"Study on structural analysis, decomposition reaction kinetics and extraction separation of green fluorescent substances in caffeine synthesis","authors":"Binbin Zhao , Jincai Yue , Bo Zhou , Lingqi Kong","doi":"10.1016/j.bej.2025.109990","DOIUrl":"10.1016/j.bej.2025.109990","url":null,"abstract":"<div><div>Caffeine is a widely used psychoactive substance. The green fluorescent substance (GFS) generated during its chemical synthesis seriously affects the appearance of products, leading to off-specification batches and increased wastewater. This study purified GFS to 95.87 % purity via preparative high-performance liquid chromatography (HPLC), identifying it as 1,3-dimethyl-7-(methylamino)-2,4-dioxo-1,2,3,4-tetrahydropteridine-6-carboxylic acid (C₁₀H₁₁N₅O₄). Quantitative analysis showed GFS forms during violuric acid hydrogenation, with partial decarboxylation in the closed-loop stage but fluctuating residues that compromise quality. Based on these findings, experimental studies were conducted on the decomposition behavior of GFS under alkaline conditions and its extraction removal characteristics. Chloroform, a polar solvent, has high extraction rate for GFS. Extraction rate increased with decreasing pH and temperature, reaching 74.92 % at pH 2 and 308.15 K when the same mass of extractant as the GFS aqueous solution was added. Decomposition reaction studies demonstrated that the decomposition rate <span><math><msub><mrow><mi>x</mi></mrow><mrow><mi>GFS</mi></mrow></msub></math></span> increased significantly with higher NaOH concentration and reaction temperature; under conditions of 8 mol·L⁻¹ NaOH, 368.15 K, and 50 min, <span><math><msub><mrow><mi>x</mi></mrow><mrow><mi>GFS</mi></mrow></msub></math></span> reached 100 %. Regression analysis yielded the following kinetic equation for the decomposition reaction under alkaline conditions: <span><math><mrow><mi>r</mi><mo>=</mo><mo>−</mo><mi>exp</mi><mo>(</mo><mfrac><mrow><mn>55.32</mn><mo>−</mo><mn>4.72</mn><mi>m</mi></mrow><mrow><mi>RT</mi></mrow></mfrac><mo>+</mo><mn>10.82</mn><mo>−</mo><mn>1.47</mn><mi>m</mi><mo>)</mo><msub><mrow><mi>C</mi></mrow><mrow><mi>A</mi></mrow></msub></mrow></math></span>. This provides critical data for a \"solvent extraction + alkaline decomposition\" process to improve caffeine production and quality, and reduce wastewater discharge.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109990"},"PeriodicalIF":3.7,"publicationDate":"2025-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Rapid lactic acid fermentation of vegetable waste in liquid-phase systems 植物废弃物液相系统快速乳酸发酵研究

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-11-01 DOI: 10.1016/j.bej.2025.109989

Han Zhang , Xue Song , Jie Zhang , Yongde Liu , Tianrun Bai , Feiyue Wang , Guihua Yan , Longjie Xiao , Tianzeng Jin , Jihong Zhao

In this study, vegetable waste liquid (VWL) was used as a substrate to produce lactic acid (LA) through anaerobic fermentation. The effects of inoculum size, carbon-to-nitrogen ratio (C/N ratio), temperature, and initial pH on LA yield and microbial succession were systematically investigated. All parameters significantly influenced LA production, with optimized conditions (12 % inoculum, initial pH of 6, C/N ratio of 20, 35 °C) yielding maximum LA production (11.76 g/L) within 16 h, corresponding to a 53.26 % glucose conversion rate. Excessive inoculum or deviation from optimal pH inhibited LA synthesis, whereas an appropriate C/N ratio enhanced microbial activity and substrate utilization. Lactobacillus plantarum was identified as the dominant species, with its relative abundance increasing from 64.14 % to 70.33 % at 16 h before stabilizing at 71.47 % at 24 h. Its metabolic dominance strongly correlated with rapid acidification, indicating its key role in efficient LA biosynthesis. Overall, precise fermentation parameter regulation effectively alleviated LA feedback inhibition and shortened the fermentation cycle to less than 24 h, offering a sustainable approach for converting vegetable waste into valuable biochemicals.

本研究以植物废液（VWL）为底物，通过厌氧发酵生产乳酸（LA）。系统研究了接种量、碳氮比（C/N）、温度和初始pH对LA产量和微生物演替的影响。所有参数均显著影响LA的产量，优化条件（接种量为12 %，初始pH为6，C/N比为20,35°C）在16 h内产生最大LA产量（11.76 g/L），对应的葡萄糖转化率为53.26% %。过量接种或偏离最佳pH会抑制LA合成，而适当的C/N比率则会增强微生物活性和底物利用率。植物乳杆菌（Lactobacillus plantarum）为优势菌种，其相对丰度在16 h时从64.14 %增加到70.33 %，在24 h时稳定在71.47 %。其代谢优势与快速酸化密切相关，表明其在高效LA生物合成中起关键作用。总体而言，精确的发酵参数调节有效地缓解了LA反馈抑制，并将发酵周期缩短至24 h以下，为将蔬菜废弃物转化为有价值的生化物质提供了一条可持续的途径。

{"title":"Rapid lactic acid fermentation of vegetable waste in liquid-phase systems","authors":"Han Zhang , Xue Song , Jie Zhang , Yongde Liu , Tianrun Bai , Feiyue Wang , Guihua Yan , Longjie Xiao , Tianzeng Jin , Jihong Zhao","doi":"10.1016/j.bej.2025.109989","DOIUrl":"10.1016/j.bej.2025.109989","url":null,"abstract":"<div><div>In this study, vegetable waste liquid (VWL) was used as a substrate to produce lactic acid (LA) through anaerobic fermentation. The effects of inoculum size, carbon-to-nitrogen ratio (C/N ratio), temperature, and initial pH on LA yield and microbial succession were systematically investigated. All parameters significantly influenced LA production, with optimized conditions (12 % inoculum, initial pH of 6, C/N ratio of 20, 35 °C) yielding maximum LA production (11.76 g/L) within 16 h, corresponding to a 53.26 % glucose conversion rate. Excessive inoculum or deviation from optimal pH inhibited LA synthesis, whereas an appropriate C/N ratio enhanced microbial activity and substrate utilization. <em>Lactobacillus plantarum</em> was identified as the dominant species, with its relative abundance increasing from 64.14 % to 70.33 % at 16 h before stabilizing at 71.47 % at 24 h. Its metabolic dominance strongly correlated with rapid acidification, indicating its key role in efficient LA biosynthesis. Overall, precise fermentation parameter regulation effectively alleviated LA feedback inhibition and shortened the fermentation cycle to less than 24 h, offering a sustainable approach for converting vegetable waste into valuable biochemicals.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109989"},"PeriodicalIF":3.7,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463346","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

STABILON enhances recombinant protein expression by mitigating proteolytic degradation in CHO cells STABILON通过减轻CHO细胞的蛋白水解降解来增强重组蛋白的表达

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-11-01 DOI: 10.1016/j.bej.2025.109988

Jia-xin Li , Ya-ru Zhao , Qiu-li Sun , Zi-meng Han , Yan-fang Wang , Xiao Guo , Xiao-yin Wang , Tian-yun Wang

Nearly 70 % of approved recombinant therapeutic proteins(RTPs) are produced in Chinese hamster ovary (CHO) cells, however, enhancing the expression levels of RTPs produced by CHO cells and mitigating their product degradation remain urgent issues to be addressed. In this study, five STABILON elements (Stab, Stab-Dm, Stab-Hs, Stab-Dm2, and Stab-Hs2) were cloned into an expression vector and transfected into CHO cells to analyze their effects on the transient expression, stable expression and proteolytic degradation of recombinant proteins. The expression levels of reporter genes and the gene of interest in CHO cells were significantly enhanced by all five STABILONs. The transient and stable expression levels of EGFP were enhanced by Stab-Hs and Stab-Dm by 1.43-fold and 2.24-fold respectively; The transient and stable expression of SEAP was elevated by Stab-Hs2 by 1.58-fold and 1.9-fold. Similarly, the stable interleukin-6 (IL-6) yield was increased by Stab-Hs2 by 1.93-fold respectively. After 30 passages of subculture, the protein retention rate of EGFP and SEAP were increased by 1.98-fold and 2.77-fold in the Stab group. Transcriptome sequencing revealed that the ubiquitin-mediated proteolytic pathway was the most significant pathway for differential gene enrichment. In conclusion, we firstly found that STABILONs can significantly enhance the transient/stable expression and recombinant protein expression by reducing proteolytic degradation during long-term passage culture in CHO cells, which is expected to be used for the industrial production of RTPs.

近70% %的已获批准的重组治疗性蛋白（RTPs）是在中国仓鼠卵巢（CHO）细胞中产生的，然而，提高CHO细胞产生的RTPs的表达水平和减轻其产物降解仍然是亟待解决的问题。本研究将5个STABILON元件（Stab、Stab- dm、Stab- hs、Stab- dm2和Stab- hs2）克隆到表达载体中，转染CHO细胞，分析其对重组蛋白瞬时表达、稳定表达和蛋白水解降解的影响。报告基因和感兴趣基因在CHO细胞中的表达水平均被这5种STABILONs显著提高。经Stab-Hs和Stab-Dm处理后，EGFP的瞬时和稳定表达水平分别提高了1.43倍和2.24倍；Stab-Hs2可使SEAP瞬时稳定表达量分别提高1.58倍和1.9倍。同样，稳定的白介素-6 （IL-6）产量通过Stab-Hs2分别提高了1.93倍。传代30代后，Stab组EGFP和SEAP的蛋白保留率分别提高了1.98倍和2.77倍。转录组测序显示，泛素介导的蛋白水解途径是差异基因富集最显著的途径。综上所述，我们首次发现，在CHO细胞的长期传代培养过程中，STABILONs可以通过减少蛋白水解降解，显著提高瞬时/稳定表达和重组蛋白的表达，有望用于rtp的工业化生产。

{"title":"STABILON enhances recombinant protein expression by mitigating proteolytic degradation in CHO cells","authors":"Jia-xin Li , Ya-ru Zhao , Qiu-li Sun , Zi-meng Han , Yan-fang Wang , Xiao Guo , Xiao-yin Wang , Tian-yun Wang","doi":"10.1016/j.bej.2025.109988","DOIUrl":"10.1016/j.bej.2025.109988","url":null,"abstract":"<div><div>Nearly 70 % of approved recombinant therapeutic proteins(RTPs) are produced in Chinese hamster ovary (CHO) cells, however, enhancing the expression levels of RTPs produced by CHO cells and mitigating their product degradation remain urgent issues to be addressed. In this study, five STABILON elements (Stab, Stab-Dm, Stab-Hs, Stab-Dm2, and Stab-Hs2) were cloned into an expression vector and transfected into CHO cells to analyze their effects on the transient expression, stable expression and proteolytic degradation of recombinant proteins. The expression levels of reporter genes and the gene of interest in CHO cells were significantly enhanced by all five STABILONs. The transient and stable expression levels of EGFP were enhanced by Stab-Hs and Stab-Dm by 1.43-fold and 2.24-fold respectively; The transient and stable expression of SEAP was elevated by Stab-Hs2 by 1.58-fold and 1.9-fold. Similarly, the stable interleukin-6 (IL-6) yield was increased by Stab-Hs2 by 1.93-fold respectively. After 30 passages of subculture, the protein retention rate of EGFP and SEAP were increased by 1.98-fold and 2.77-fold in the Stab group. Transcriptome sequencing revealed that the ubiquitin-mediated proteolytic pathway was the most significant pathway for differential gene enrichment. In conclusion, we firstly found that STABILONs can significantly enhance the transient/stable expression and recombinant protein expression by reducing proteolytic degradation during long-term passage culture in CHO cells, which is expected to be used for the industrial production of RTPs.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109988"},"PeriodicalIF":3.7,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Treatment of real aquaculture wastewater using marine Chlorella: Pollutant removal and microbial community dynamics 用海洋小球藻处理真实水产养殖废水：污染物去除和微生物群落动态

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-31 DOI: 10.1016/j.bej.2025.109987

Shenwei Cheng , Yanqing Sheng

This study comprehensively investigated the nutrient removal mechanisms of Chlorella sp. in authentic aquaculture wastewater, while also examining the dynamic shifts within its symbiotic microbial community. The results showed that the microalgae exhibited robust growth in the wastewater, effectively reducing concentrations of ammonium nitrogen, nitrate nitrogen, and phosphate with removal efficiencies of 95.0 %, 93.2 %, and 91.7 %, respectively. Fluctuations in pH during the initial cultivation period indicated a cyclical interplay between heterotrophic microbial activity and microalgal photosynthesis. In the later stages, enhanced synthesis of Chlorella carotenoids was correlated with deteriorating water quality and the onset of oxidative stress. Initially, the microbial community was dominated by organic matter degradation, nitrate reduction, and fermentation. However, as cultivation progressed, metabolic activities shifted towards the photoassimilation of inorganic nutrients. The Chlorella sp. symbiotic system facilitated a rapid succession of the microbial community, which was characterized by frequent bacterial population changes, ultimately leading to a unique, temporally stable community structure. The initially diverse microbial population was progressively supplanted by more adaptive bacterial strains, achieving a steady state by the seventh day. Notably, Alcaligenaceae demonstrated exceptional adaptability compared to other genera, underscoring its pivotal role within the Chlorella sp. symbiotic system. This research provides valuable theoretical insights into algal-bacterial interactions and highlights their potential for application in water quality remediation.

本研究全面探讨了小球藻在真实养殖废水中的营养去除机制，同时考察了其共生微生物群落的动态变化。结果表明，微藻在废水中生长旺盛，能有效降低铵态氮、硝态氮和磷酸盐的浓度，去除率分别为95.0 %、93.2 %和91.7 %。培养初期pH值的波动表明异养微生物活性与微藻光合作用之间存在周期性相互作用。在后期，小球藻类胡萝卜素合成的增加与水质恶化和氧化应激的发生有关。最初，微生物群落以有机物降解、硝酸盐还原和发酵为主。然而，随着栽培的进行，代谢活动转向无机养分的光同化。小球藻共生系统促进了微生物群落的快速演替，其特征是细菌种群的频繁变化，最终形成了一个独特的、暂时稳定的群落结构。最初多样化的微生物种群逐渐被适应性更强的菌株所取代，在第7天达到稳定状态。值得注意的是，与其他属相比，Alcaligenaceae表现出特殊的适应性，强调了它在小球藻共生系统中的关键作用。该研究为藻-细菌相互作用提供了有价值的理论见解，并强调了它们在水质修复中的应用潜力。

{"title":"Treatment of real aquaculture wastewater using marine Chlorella: Pollutant removal and microbial community dynamics","authors":"Shenwei Cheng , Yanqing Sheng","doi":"10.1016/j.bej.2025.109987","DOIUrl":"10.1016/j.bej.2025.109987","url":null,"abstract":"<div><div>This study comprehensively investigated the nutrient removal mechanisms of <em>Chlorella</em> sp. in authentic aquaculture wastewater, while also examining the dynamic shifts within its symbiotic microbial community. The results showed that the microalgae exhibited robust growth in the wastewater, effectively reducing concentrations of ammonium nitrogen, nitrate nitrogen, and phosphate with removal efficiencies of 95.0 %, 93.2 %, and 91.7 %, respectively. Fluctuations in pH during the initial cultivation period indicated a cyclical interplay between heterotrophic microbial activity and microalgal photosynthesis. In the later stages, enhanced synthesis of <em>Chlorella</em> carotenoids was correlated with deteriorating water quality and the onset of oxidative stress. Initially, the microbial community was dominated by organic matter degradation, nitrate reduction, and fermentation. However, as cultivation progressed, metabolic activities shifted towards the photoassimilation of inorganic nutrients. The <em>Chlorella</em> sp. symbiotic system facilitated a rapid succession of the microbial community, which was characterized by frequent bacterial population changes, ultimately leading to a unique, temporally stable community structure. The initially diverse microbial population was progressively supplanted by more adaptive bacterial strains, achieving a steady state by the seventh day. Notably, <em>Alcaligenaceae</em> demonstrated exceptional adaptability compared to other genera, underscoring its pivotal role within the <em>Chlorella</em> sp. symbiotic system. This research provides valuable theoretical insights into algal-bacterial interactions and highlights their potential for application in water quality remediation.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109987"},"PeriodicalIF":3.7,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Temperature-tailored cellulose-rich Cladophora sp. hydrochar for selective malachite green adsorption from multi-dye systems 温度定制的富含纤维素的Cladophora sp.碳氢化合物对多染料系统中孔雀石绿的选择性吸附

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-30 DOI: 10.1016/j.bej.2025.109986

Muhammad Badaruddin , Laila Hanum , Elda Melwita , Sahrul Wibiyan , Yulizah Hanifah , Aldes Lesbani

This study presents the selective adsorption of malachite green (MG) from multi-dye mixtures using hydrochar derived from Cladophora sp. (C) via hydrothermal carbonization (HTC) at 150°C (C150), 200°C (C200), 250°C (C250), and 300°C (C300). Structural characterizations (XRD, FTIR, SEM-EDS, BET) confirmed enhanced porosity and surface functionalization with increasing HTC temperature. C200 exhibited the highest BET surface area (15.84 m²/g), while C300, with the largest pore size (18.27 nm), showed the highest MG adsorption capacity (204.1 mg/g), outperforming C (30.03 mg/g). Selective adsorption tests using ternary anionic-cationic dye mixtures revealed strong affinity toward cationic dyes, with MG showing 95.5 % removal by C300. Kinetic studies fitted the pseudo-second-order model, suggesting chemisorption dominance; the Avrami model also correlated well. Increasing Avrami rate constant (k₃) and decreasing exponent (n) with HTC temperature indicate a transition from diffusion to surface-controlled chemisorption. Equilibrium data followed the Sips isotherm, reflecting mixed monolayer–multilayer adsorption on moderately heterogeneous surfaces. The rising Sips constant (Kₛ) with temperature confirmed an endothermic process consistent with thermodynamic findings, which also verified spontaneity. Regeneration studies showed C300 retained > 50 % efficiency after three cycles, demonstrating stability and reusability. FTIR, XRD, and XPS analyses before and after MG adsorption confirmed interactions via electrostatic attraction, hydrogen bonding, and π–π interactions.

本研究介绍了从Cladophora sp. (C)衍生的碳氢化合物（HTC）在150°C (C150), 200°C (C200), 250°C （C250）和300°C （C300）下通过水热碳化（HTC）从多种染料混合物中选择性吸附孔雀石绿（MG）。结构表征（XRD， FTIR, SEM-EDS， BET）证实，随着HTC温度的升高，孔隙率和表面功能化增强。C200的BET表面积最大（15.84 m²/g）， C300的吸附量最大（18.27 nm），达到204.1 MG /g，优于C（30.03 MG /g）。使用三元阴离子-阳离子染料混合物的选择性吸附试验表明，对阳离子染料有很强的亲和力，C300对MG的去除率为95.5 %。动力学研究符合伪二阶模型，表明化学吸附优势；Avrami模型也有很好的相关性。Avrami速率常数（k₃）随着HTC温度的升高而增大，指数(n)随着HTC温度的升高而减小，表明由扩散向表面控制的化学吸附转变。平衡数据遵循Sips等温线，反映了在中等非均质表面上的混合单层-多层吸附。Sips常数（Kₛ）随温度的升高证实了一个与热力学结果一致的吸热过程，这也证实了自发性。再生研究表明，C300在三次循环后保持了>； 50 %的效率，表现出稳定性和可重复使用性。吸附MG前后的FTIR、XRD和XPS分析证实了静电吸引、氢键和π -π相互作用。

{"title":"Temperature-tailored cellulose-rich Cladophora sp. hydrochar for selective malachite green adsorption from multi-dye systems","authors":"Muhammad Badaruddin , Laila Hanum , Elda Melwita , Sahrul Wibiyan , Yulizah Hanifah , Aldes Lesbani","doi":"10.1016/j.bej.2025.109986","DOIUrl":"10.1016/j.bej.2025.109986","url":null,"abstract":"<div><div>This study presents the selective adsorption of malachite green (MG) from multi-dye mixtures using hydrochar derived from <em>Cladophora</em> sp<em>.</em> (C) via hydrothermal carbonization (HTC) at 150°C (C150), 200°C (C200), 250°C (C250), and 300°C (C300). Structural characterizations (XRD, FTIR, SEM-EDS, BET) confirmed enhanced porosity and surface functionalization with increasing HTC temperature. C200 exhibited the highest BET surface area (15.84 m²/g), while C300, with the largest pore size (18.27 nm), showed the highest MG adsorption capacity (204.1 mg/g), outperforming C (30.03 mg/g). Selective adsorption tests using ternary anionic-cationic dye mixtures revealed strong affinity toward cationic dyes, with MG showing 95.5 % removal by C300. Kinetic studies fitted the pseudo-second-order model, suggesting chemisorption dominance; the Avrami model also correlated well. Increasing Avrami rate constant (k₃) and decreasing exponent (n) with HTC temperature indicate a transition from diffusion to surface-controlled chemisorption. Equilibrium data followed the Sips isotherm, reflecting mixed monolayer–multilayer adsorption on moderately heterogeneous surfaces. The rising Sips constant (Kₛ) with temperature confirmed an endothermic process consistent with thermodynamic findings, which also verified spontaneity. Regeneration studies showed C300 retained > 50 % efficiency after three cycles, demonstrating stability and reusability. FTIR, XRD, and XPS analyses before and after MG adsorption confirmed interactions via electrostatic attraction, hydrogen bonding, and π–π interactions.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109986"},"PeriodicalIF":3.7,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145463344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Metabolic coordination of ammonium assimilation and denitrification enhances nitrogen removal by Rhodococcus biphenylivorans XYH-1 in wastewater treatment 氨同化和反硝化的代谢协调促进了联苯利华红球菌XYH-1在废水处理中的脱氮作用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-30 DOI: 10.1016/j.bej.2025.109985

Yujia Li , Hao Xu , Dongyun Ye , Hongye Fan , Dzmitry Hrynsphan , Savitskaya Tatsiana , Zeyu Wang , Jun Chen

Ammonium and nitrate frequently coexist in wastewater, yet the metabolic coordination between their removal remains unclear. The potential trade-off between ammonium assimilation and denitrification limits the efficiency of biological treatment. This study investigated nitrogen transformation in Rhodococcus biphenylivorans XYH-1 under mixed nitrogen conditions. At 50 mg·L⁻¹ NH₄⁺-N and 100 mg·L⁻¹ NO₃^--N, total nitrogen (TN) removal reached 82.3 %, with denitrification and assimilation contributing 43.7 % and 38.6 %, respectively. Ammonium assimilation upregulated napA (2.36-fold), nasR (2.25-fold), and gltB (1.91-fold), promoting biomass synthesis (0.112 g·L⁻¹) and NO₃^--N reduction. However, at 100 mg·L⁻¹ NH₄⁺-N, the expression of norB and nosZ expression decreased by 37.2 % and 40.5 %, respectively, resulting in NO₂^--N accumulation (6.41 mg·L⁻¹) and a decline in TN removal to 58.6 %. This concentration-dependent shift highlights that moderate NH₄⁺-N enhances nitrogen removal via coordinated metabolic routing, whereas excess NH₄⁺-N disrupts electron flow and enzyme expression. These findings provide mechanistic insight for optimizing nitrogen-rich wastewater treatment.

铵态氮和硝态氮在废水中经常共存，但它们之间的代谢协调作用尚不清楚。氨同化和反硝化之间的潜在权衡限制了生物处理的效率。本研究研究了在混合氮条件下，联苯livorans红球菌XYH-1的氮转化。在50 mg·L−1 NH4+-N和100 mg·L−1 NO3——N条件下，总氮去除率达到82.3 %，其中反硝化作用和同化作用分别贡献43.7% %和38.6% %。铵同化上调napA（2.36倍）、nasR（2.25倍）和gltB（1.91倍），促进生物质合成（0.112 g·L−1）和NO3—N还原。而当NH4+-N浓度为100 mg·L−1时，norB和nosZ的表达量分别下降37.2% %和40.5 %，NO2——N积累量下降6.41 mg·L−1，TN去除率下降58.6% %。这种浓度依赖性的转变表明，适度的NH4+-N通过协调的代谢途径促进氮的去除，而过量的NH4+-N会破坏电子流和酶的表达。这些发现为优化富氮废水处理提供了机理见解。

{"title":"Metabolic coordination of ammonium assimilation and denitrification enhances nitrogen removal by Rhodococcus biphenylivorans XYH-1 in wastewater treatment","authors":"Yujia Li , Hao Xu , Dongyun Ye , Hongye Fan , Dzmitry Hrynsphan , Savitskaya Tatsiana , Zeyu Wang , Jun Chen","doi":"10.1016/j.bej.2025.109985","DOIUrl":"10.1016/j.bej.2025.109985","url":null,"abstract":"<div><div>Ammonium and nitrate frequently coexist in wastewater, yet the metabolic coordination between their removal remains unclear. The potential trade-off between ammonium assimilation and denitrification limits the efficiency of biological treatment. This study investigated nitrogen transformation in <em>Rhodococcus biphenylivorans</em> XYH-1 under mixed nitrogen conditions. At 50 mg·L<sup>−1</sup> NH<sub>4</sub><sup>+</sup>-N and 100 mg·L<sup>−1</sup> NO<sub>3</sub><sup>-</sup>-N, total nitrogen (TN) removal reached 82.3 %, with denitrification and assimilation contributing 43.7 % and 38.6 %, respectively. Ammonium assimilation upregulated <em>napA</em> (2.36-fold), <em>nasR</em> (2.25-fold), and <em>gltB</em> (1.91-fold), promoting biomass synthesis (0.112 g·L<sup>−1</sup>) and NO<sub>3</sub><sup>-</sup>-N reduction. However, at 100 mg·L<sup>−1</sup> NH<sub>4</sub><sup>+</sup>-N, the expression of <em>norB</em> and <em>nosZ</em> expression decreased by 37.2 % and 40.5 %, respectively, resulting in NO<sub>2</sub><sup>-</sup>-N accumulation (6.41 mg·L<sup>−1</sup>) and a decline in TN removal to 58.6 %. This concentration-dependent shift highlights that moderate NH<sub>4</sub><sup>+</sup>-N enhances nitrogen removal via coordinated metabolic routing, whereas excess NH<sub>4</sub><sup>+</sup>-N disrupts electron flow and enzyme expression. These findings provide mechanistic insight for optimizing nitrogen-rich wastewater treatment.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109985"},"PeriodicalIF":3.7,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145413343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Machine learning optimization of pullulan production from banana peel extract by Aureobasidium pullulans 香蕉皮提取物普鲁兰毛霉生产普鲁兰的机器学习优化

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-24 DOI: 10.1016/j.bej.2025.109984

M. Pradeep , Nageswar Sahu , Biswanath Mahanty

Valorization of fruit processing waste to value-added compounds offers a sustainable microbial bioprocess. In this study, an increasing proportion of pretreated and untreated banana peel extract (BPE) was used to replace sucrose in pullulan production by Aureobasidium pullulans. The pullulan production (33.66 ± 0.57 g L⁻¹) with pretreated BPE (40 g L⁻¹ sucrose equivalent) was significantly higher than that in pure sucrose (8.33 ± 2.30 g L⁻¹) or untreated BPE sets (26.66 ± 1.52 g L⁻¹). Bayesian hyperparameter-optimized ML model variants, i.e., support vector regression, artificial neural networks, least-squares boosting, and random forest (RF) models, were developed from Gaussian noise-augmented experimental data for biopolymer and biomass production. The RF model exhibited the best performance for both pullulan (R²: 0.992, R²_CV: 0.977) and biomass (R²: 0.939, R²_CV: 0.891). Explainable AI tools revealed that initial biomass, BPE concentration, and pretreatment status were key influencers of the process. Genetic algorithm-based multi-objective optimization offered an array of Pareto-optimal solutions. This study presents a scalable strategy for low-cost, sustainable production of pullulan from agro-industrial residues.

水果加工废弃物转化为增值化合物提供了一种可持续的微生物过程。本研究通过增加预处理和未处理香蕉皮提取物（BPE）的比例来替代普鲁兰小孢子菌生产普鲁兰所需的蔗糖。支链淀粉生产(33.66 ±0.57  g L−1)与BPE预处理(40 g L−1蔗糖当量)明显高于纯蔗糖(8.33 ±2.30  g L−1)或未经处理的BPE集(26.66 ±1.52  g L−1)。贝叶斯超参数优化的ML模型变体，即支持向量回归、人工神经网络、最小二乘增强和随机森林（RF）模型，是根据生物聚合物和生物质生产的高斯噪声增强实验数据开发的。该模型对普鲁兰（R2: 0.992, R2CV: 0.977）和生物量（R2: 0.939, R2CV: 0.891）均表现最佳。可解释的AI工具显示，初始生物量、BPE浓度和预处理状态是该过程的关键影响因素。基于遗传算法的多目标优化提供了一组pareto最优解。本研究提出了一种可扩展的策略，以低成本、可持续的方式从农业工业残留物中生产普鲁兰。

{"title":"Machine learning optimization of pullulan production from banana peel extract by Aureobasidium pullulans","authors":"M. Pradeep , Nageswar Sahu , Biswanath Mahanty","doi":"10.1016/j.bej.2025.109984","DOIUrl":"10.1016/j.bej.2025.109984","url":null,"abstract":"<div><div>Valorization of fruit processing waste to value-added compounds offers a sustainable microbial bioprocess. In this study, an increasing proportion of pretreated and untreated banana peel extract (BPE) was used to replace sucrose in pullulan production by <em>Aureobasidium pullulans</em>. The pullulan production (33.66 ± 0.57 g L<sup>−1</sup>) with pretreated BPE (40 g L<sup>−1</sup> sucrose equivalent) was significantly higher than that in pure sucrose (8.33 ± 2.30 g L<sup>−1</sup>) or untreated BPE sets (26.66 ± 1.52 g L<sup>−1</sup>). Bayesian hyperparameter-optimized ML model variants, i.e., support vector regression, artificial neural networks, least-squares boosting, and random forest (RF) models, were developed from Gaussian noise-augmented experimental data for biopolymer and biomass production. The RF model exhibited the best performance for both pullulan (R<sup>2</sup>: 0.992, R<sup>2</sup><sub>CV</sub>: 0.977) and biomass (R<sup>2</sup>: 0.939, R<sup>2</sup><sub>CV</sub>: 0.891). Explainable AI tools revealed that initial biomass, BPE concentration, and pretreatment status were key influencers of the process. Genetic algorithm-based multi-objective optimization offered an array of Pareto-optimal solutions. This study presents a scalable strategy for low-cost, sustainable production of pullulan from agro-industrial residues.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109984"},"PeriodicalIF":3.7,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145413344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Kinetic and diffusional modeling of isoniazid adsorption onto biopolymer beads for organic-solvent-based separation 异烟肼吸附在生物聚合物微球上的动力学和扩散模型

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-22 DOI: 10.1016/j.bej.2025.109981

W.S. de Freitas , R.C. Jacome , I.R.F. Rocha , F.D.A. da Costa , L.R.B. Gonçalves , W.S. Adriano

The control of isoniazid (INH) in pharmaceutical and environmental contexts requires cost-effective adsorbents capable of operating under non-aqueous conditions. This study investigates the adsorption of INH onto glutaraldehyde-crosslinked chitosan beads in isopropanol. Adsorption isotherms indicated a maximum adsorption capacity of 24.37 mg.g⁻¹ at 298 K (Langmuir model), while Dubinin–Radushkevich analysis yielded an adsorption energy of 1.03 kJ.mol⁻¹ , consistent with physisorption. Kinetic models (pseudo-homogeneous, hybrid), supported by Weber–Morris, Boyd, and Horstmann – Chase analyses, and revealed intraparticle diffusion as the main resistance. Thermodynamic parameters confirmed that adsorption is spontaneous (ΔG < 0) and entropy-driven. Beyond mechanistic insights, adsorption offers a cost-effective alternative to more sophisticated separation methods, with multiple models supporting the reliability of the interpretation and significant performance maintained in isopropanol, confirming robustness under restrictive solvent conditions. Comparative analysis with literature shows that higher capacities are reported in aqueous systems, while data in organic-rich media remain scarce. These results provide a mechanistic basis for extending adsorption studies of pharmaceutical compounds to solvent-rich environments and indicate future prospects for reusability and scale-up.

在制药和环境环境中控制异烟肼（INH）需要能够在非水条件下操作的具有成本效益的吸附剂。研究了戊二醛交联壳聚糖珠在异丙醇中的吸附作用。吸附等温线显示其最大吸附量为24.37 mg。g⁻¹ 在298 K （Langmuir模型），而Dubinin-Radushkevich分析得出的吸附能为1.03 kJ。Mol⁻¹ ，与物理吸收相一致。动力学模型（伪均匀、混合）得到Weber-Morris、Boyd和Horstmann - Chase分析的支持，并揭示了颗粒内扩散是主要阻力。热力学参数证实吸附是自发的（ΔG < 0）和熵驱动的。除了对机理的深入了解之外，吸附法为更复杂的分离方法提供了一种经济有效的替代方法，多种模型支持解释的可靠性，并且在异丙醇中保持了显著的性能，证实了在限制性溶剂条件下的稳健性。与文献的比较分析表明，在水系统中有较高的容量，而在富有机介质中的数据仍然很少。这些结果为将药物化合物的吸附研究扩展到富溶剂环境提供了机制基础，并表明了可重复使用和扩大规模的未来前景。

{"title":"Kinetic and diffusional modeling of isoniazid adsorption onto biopolymer beads for organic-solvent-based separation","authors":"W.S. de Freitas , R.C. Jacome , I.R.F. Rocha , F.D.A. da Costa , L.R.B. Gonçalves , W.S. Adriano","doi":"10.1016/j.bej.2025.109981","DOIUrl":"10.1016/j.bej.2025.109981","url":null,"abstract":"<div><div>The control of isoniazid (INH) in pharmaceutical and environmental contexts requires cost-effective adsorbents capable of operating under non-aqueous conditions. This study investigates the adsorption of INH onto glutaraldehyde-crosslinked chitosan beads in isopropanol. Adsorption isotherms indicated a maximum adsorption capacity of 24.37 mg.g⁻¹ at 298 K (Langmuir model), while Dubinin–Radushkevich analysis yielded an adsorption energy of 1.03 kJ.mol⁻¹ , consistent with physisorption. Kinetic models (pseudo-homogeneous, hybrid), supported by Weber–Morris, Boyd, and Horstmann – Chase analyses, and revealed intraparticle diffusion as the main resistance. Thermodynamic parameters confirmed that adsorption is spontaneous (ΔG < 0) and entropy-driven. Beyond mechanistic insights, adsorption offers a cost-effective alternative to more sophisticated separation methods, with multiple models supporting the reliability of the interpretation and significant performance maintained in isopropanol, confirming robustness under restrictive solvent conditions. Comparative analysis with literature shows that higher capacities are reported in aqueous systems, while data in organic-rich media remain scarce. These results provide a mechanistic basis for extending adsorption studies of pharmaceutical compounds to solvent-rich environments and indicate future prospects for reusability and scale-up.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109981"},"PeriodicalIF":3.7,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145359377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In situ directed evolution of γ-glutamyl kinase in Corynebacterium glutamicum S9114 by innovative EvolvR-mediated mutagenesis and rare codon-based selection to increase L-proline production 利用创新性evolvr介导的诱变和基于罕见密码子的选择，谷氨酸棒状杆菌S9114的γ-谷氨酰基激酶原位定向进化以增加l -脯氨酸的产量

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2025-10-17 DOI: 10.1016/j.bej.2025.109982

Jian-Zhong Xu , Feng Zhang , Kai Wang , Shuo Wan , Wei-Guo Zhang

An efficient evolution and selection system is vital for constructing the high added-value products hyper-producers in fermentation industry. Herein, an innovative EvolvR-mediated mutagenesis was used to diversify the γ-glutamyl kinase (GK)-coding gene proB in C. glutamicum S9114 and an optimized rare codon-based selection was used to isolate the L-proline high-yielding strain with optimal GK variant. Triple-mutant GK^{E74K/G149K/V150M} was identified with high substrate affinity and catalytic efficiency for producing L-proline. The resultant strain C. glutamicum Pro-7 produced 94.2 ± 5.6 g/L of L-proline with productivity of 1.96 g/(L·h) and a yield of 0.316 g/g. Subsequently, the molecular mechanisms involved in high L-proline production in strain Pro-7 was discovered, indicating that ATP and NADPH supply are also the key factors for the L-proline biosynthesis besides GK. Thus, in situ directed evolution of key genes in genome by EvolvR-mediated mutagenesis and rare codon-based selection provides a new direction and reference to construct industrial-strength strains.

高效的进化和选择系统是构建发酵工业高附加值产品超级生产者的关键。本文采用创新性的evolvr介导诱变技术，对C. glutamicum S9114中γ-谷氨酰基激酶（GK）编码基因probb进行了突变，并采用优化的稀有密码子选择方法分离出了具有最佳GK变异的l -脯氨酸高产菌株。三突变体GKE74K/G149K/V150M对l -脯氨酸具有较高的底物亲和力和催化效率。菌株C. glutamum Pro-7的L-脯氨酸产量为94.2 ± 5.6 g/L，产率为1.96 g/(L·h)，产率为0.316 g/g。随后发现菌株Pro-7高产l -脯氨酸的分子机制，表明除了GK外，ATP和NADPH的供应也是l -脯氨酸生物合成的关键因素。因此，通过evolvr介导的突变和基于稀有密码子的选择对基因组关键基因进行原位定向进化，为构建工业强度菌株提供了新的方向和参考。

{"title":"In situ directed evolution of γ-glutamyl kinase in Corynebacterium glutamicum S9114 by innovative EvolvR-mediated mutagenesis and rare codon-based selection to increase L-proline production","authors":"Jian-Zhong Xu , Feng Zhang , Kai Wang , Shuo Wan , Wei-Guo Zhang","doi":"10.1016/j.bej.2025.109982","DOIUrl":"10.1016/j.bej.2025.109982","url":null,"abstract":"<div><div>An efficient evolution and selection system is vital for constructing the high added-value products hyper-producers in fermentation industry. Herein, an innovative EvolvR-mediated mutagenesis was used to diversify the γ-glutamyl kinase (GK)-coding gene <em>proB</em> in <em>C. glutamicum</em> S9114 and an optimized rare codon-based selection was used to isolate the L-proline high-yielding strain with optimal GK variant. Triple-mutant GK<sup>E74K/G149K/V150M</sup> was identified with high substrate affinity and catalytic efficiency for producing L-proline. The resultant strain <em>C. glutamicum</em> Pro-7 produced 94.2 ± 5.6 g/L of L-proline with productivity of 1.96 g/(L·h) and a yield of 0.316 g/g. Subsequently, the molecular mechanisms involved in high L-proline production in strain Pro-7 was discovered, indicating that ATP and NADPH supply are also the key factors for the L-proline biosynthesis besides GK. Thus, <em>in situ</em> directed evolution of key genes in genome by EvolvR-mediated mutagenesis and rare codon-based selection provides a new direction and reference to construct industrial-strength strains.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"226 ","pages":"Article 109982"},"PeriodicalIF":3.7,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145326137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0