Biochemical Engineering Journal最新文献_第5页

Whole-cell synthesis of nicotinamide mononucleotide by recombinant Saccharomyces cerevisiae from glucose and nicotinamide 重组酿酒酵母从葡萄糖和烟酰胺全细胞合成烟酰胺单核苷酸

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-10 DOI: 10.1016/j.bej.2024.109528

Chaoguang Wang , Xiaohan Hui , George Marshall , Wenhan Xiao , Xiaomei Zhang , Jianying Qian , Jinsong Gong , Guoqiang Xu , Jinsong Shi , Zhenghong Xu

β-Nicotinamide mononucleotide is recognized as a significant bioactive nucleotide, which is can be used in the fields of health industries. Many studies on the synthesis of NMN have involved Escherichia coli and the current methods are limited by safety problems as well as the expense of the substrate. Herein, GRAS-grade Saccharomyces cerevisiae was chosen as chassis cells to synthesize NMN using the substrates glucose and nicotinamide. First, the gene for the key enzyme nicotinamide phosphoribosyltransferase (Nampt) was screened from different sources, and site-directed mutation was performed to improve the synthesis of NMN. The concentration of intracellular NMN in yeast expressing the D83N-Nampt mutant derived from Chitinophaga pinensis reached 413.4 mg/L, which was 3.7 times higher than that of yeast expressing wild enzymes. The synthesis of NMN was further enhanced by overexpressing Nampt combined with weakening of the further metabolism of NMN. Subsequently, the supply of precursor phosphate ribose pyrophosphate (PRPP) was increased by overexpressing the PRPP synthase mutant, which led to the concentration of intracellular NMN increased to 775.9 mg/L from 537.8 mg/L. Finally, the concentration of intracellular NMN reached 1.2 g/L at 6 h after whole-cell catalytic optimization, which is the highest titer achieved by S. cerevisiae from inexpensive substrate glucose and nicotinamide. This study provides the synthesis of NMN by S. cerevisiae with a new and promising method.

β-烟酰胺单核苷酸被认为是一种重要的生物活性核苷酸，可用于健康产业领域。许多关于合成 NMN 的研究都涉及到大肠杆菌，目前的方法受到安全问题和底物昂贵的限制。本文选择 GRAS 级的酿酒酵母作为底盘细胞，以葡萄糖和烟酰胺为底物合成 NMN。首先，从不同来源筛选出关键酶烟酰胺磷酸核糖转移酶（Nampt）的基因，并进行定点突变以提高 NMN 的合成。在表达来自 Chitinophaga pinensis 的 D83N-Nampt 突变体的酵母中，细胞内 NMN 的浓度达到 413.4 mg/L，是表达野生酶的酵母的 3.7 倍。过表达 Nampt 进一步提高了 NMN 的合成，同时削弱了 NMN 的进一步代谢。随后，通过过表达 PRPP 合成酶突变体，增加了前体磷酸核糖焦磷酸（PRPP）的供应，从而使细胞内 NMN 的浓度从 537.8 mg/L 增加到 775.9 mg/L。最后，经过全细胞催化优化后，细胞内 NMN 的浓度在 6 h 时达到 1.2 g/L，这是 S. cerevisiae 从廉价底物葡萄糖和烟酰胺中获得的最高滴度。这项研究为利用酿酒酵母合成 NMN 提供了一种新的可行方法。

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引用次数: 0

Process development and environmental impact assessment of sustainable poly(3-hydroxybutyrate) separation and purification via Paraburkholderia sacchari cell lysis using crude enzymes 利用粗酶通过 Paraburkholderia sacchari 细胞裂解分离和纯化可持续聚（3-羟基丁酸）的工艺开发和环境影响评估

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-10 DOI: 10.1016/j.bej.2024.109526

Olga Psaki , Lina Zoghbi , Drosoula Despoina Galani , Anastasios Giannoulis , Ioanna-Georgia I. Athanasoulia , Eleni Stylianou , Demetres Briassoulis , Apostolis Koutinas , Dimitrios Ladakis

Downstream separation and purification of poly(3-hydroxybutyrate) has been developed via combined crude enzyme bacterial cell lysis followed by solvent extraction and circular utilisation of bacterial lysate as nutrient supplement for PHB production. Enzymatic lysis of Paraburkholderia sacchari cells was initially evaluated using crude enzymes produced via solid state fermentation of Aspergillus oryzae. The optimum protease activity (156 U per g cell dry weight) resulted in 66.9 % lysis of residual cell weight. PHB purification with 1,3-dioxolane, dimethyl carbonate, anisole, ammonium laurate and chloroform were evaluated at different processing parameters (extraction duration, temperature) using wet and dry bacterial cells as well as enzymatically lysed wet bacterial cells. The highest recovery yield (94.5 %) and purity (98.1 %) were obtained with 1,3-dioxolane at 80°C and 6 h processing time using enzymatically disrupted bacterial cells. The bacterial cell lysate was used as nutrient supplement for circular PHB production in fed-batch P. sacchari bioreactor culture leading to the production of 78.7 g_PHB/L, 56.9 % (w/w) PHB content and 2.3 g/(Lꞏh) productivity. Low global warming potential (1.3 CO₂-eq/kg_PHB) and abiotic depletion fossil (14.4 MJ/kg_PHB) were estimated for PHB purification via enzymatic cell lysis and PHB extraction with 1,3-dioxolane demonstrating the development of a sustainable and circular process for PHB production.

聚（3-羟基丁酸）的下游分离和纯化是通过粗酶细菌细胞裂解，然后进行溶剂萃取和循环利用细菌裂解物作为 PHB 生产的营养补充剂而实现的。最初使用通过黑曲霉（Aspergillus oryzae）固态发酵产生的粗酶对 Paraburkholderia sacchari 细胞的酶裂解进行了评估。最佳蛋白酶活性（每克细胞干重 156 U）可使残余细胞重量的 66.9% 降解。在不同的处理参数（萃取持续时间、温度）下，使用干湿细菌细胞以及酶裂解湿细菌细胞，评估了用 1,3-二氧戊环、碳酸二甲酯、苯甲醚、月桂酸铵和氯仿提纯 PHB 的效果。在 80°C 和 6 小时处理时间内，使用酶解细菌细胞萃取 1,3-二氧戊环的回收率（94.5%）和纯度（98.1%）最高。将细菌细胞裂解液作为营养补充物，在喂料批次 P. sacchari 生物反应器培养中进行循环 PHB 生产，可获得 78.7 gPHB/L、56.9 %（w/w）PHB 含量和 2.3 g/(Lꞏh) 生产率。通过酶解细胞和用 1,3-二氧戊环提取 PHB 来纯化 PHB，估计全球变暖潜能值（1.3 CO2-eq/kgPHB）和非生物损耗化石（14.4 MJ/kgPHB）较低，这表明开发出了一种可持续和循环的 PHB 生产工艺。

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引用次数: 0

Insights into Pb (II) adsorption mechanisms using jackfruit peel biochar activated by a hydrothermal method toward heavy metal removal from wastewater 利用水热法活化的菠萝皮生物炭去除废水中重金属的铅 (II) 吸附机理探析

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-10 DOI: 10.1016/j.bej.2024.109525

Loc Ton-That , Thi-Phuong-Tu Nguyen , Bich-Ngoc Duong , Duy-Khoi Nguyen , Ngoc-An Nguyen , Thien‑Hoang Ho , Van-Phuc Dinh

Understanding adsorption mechanisms plays an instrumental role in designing and operating adsorption-based wastewater treatment systems. This research systematically demonstrated the comprehensive adsorption mechanism of Pb(II) ions onto biochar synthesized from jackfruit peel in an aqueous solution using theoretical adsorption models and cutting-edge analytical techniques, such as FT-IR, TG-DSC, and SEM-EDX. The results showed that the adsorption process followed the Redlich-Peterson isothermal model and the intraparticle diffusion kinetic model under optimized conditions. The key mechanisms contributing to effective Pb(II) adsorption include complexation, ion - exchange, and intradiffusion. Furthermore, using the hydrothermal method to active biochar improves the surface area of JPT, leading to the maximum Pb(II) adsorption capacity of jackfruit peel-derived biochar to be 83.86 mg/g, higher than biochars from other parts of jackfruit waste (seeds and stems) and some materials from different agricultural residues in previous studies. These findings contribute to narrowing the gap in understanding heavy metal adsorption using biomass residues, theoretical models, and their mechanisms. Additionally, these indicate that agricultural by-products such as jackfruit peel are environmentally friendly and economical materials for implementing strategies aimed at mitigating heavy metal pollution in wastewater.

了解吸附机理对设计和运行基于吸附的废水处理系统具有重要作用。本研究利用理论吸附模型和 FT-IR、TG-DSC 和 SEM-EDX 等前沿分析技术，系统地论证了水溶液中铅(II)离子在由柚子皮合成的生物炭上的综合吸附机理。结果表明，在优化条件下，吸附过程遵循 Redlich-Peterson 等温模型和颗粒内扩散动力学模型。有效吸附铅（II）的关键机制包括络合、离子交换和内扩散。此外，水热法提高了活性生物炭的表面积，从而使柚子皮衍生生物炭的最大铅（II）吸附容量达到 83.86 mg/g，高于以往研究中柚子废物其他部分（种子和茎）的生物炭以及来自不同农业残留物的一些材料。这些发现有助于缩小对生物质残渣重金属吸附、理论模型及其机理的认识差距。此外，这些研究还表明，柚子皮等农副产品是实施旨在减轻废水中重金属污染的策略的环保而经济的材料。

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引用次数: 0

Characterization of the dynamic viscosity of cell cultures and its effect on mixing performance in a spinner flask bioreactor 细胞培养物动态粘度的特征及其对旋转瓶生物反应器混合性能的影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-09 DOI: 10.1016/j.bej.2024.109523

Fernando Cantarero-Rivera , Doris H. D’Souza , Madhu Dhar , Jiajia Chen

Computational fluid dynamics (CFD) models have been developed to simulate cell culturing bioreactors but assume water-like viscosity properties due to significant data gaps. This study characterized the dynamic viscosity of HEK-293 cell cultures and evaluated its effect on mixing performance in a spinner flask bioreactor. Viscosity measurements indicated that the cell culture media, media with microcarriers, and cell cultures presented shear thinning behaviors within the measured shear rate range of 1 to 100 s^-1. The viscosity also increased with the microcarrier concentrations and growth of cell culture. The CFD model, incorporating dynamic viscosity data, showed that shear stress and Kolmogorov length profiles are significantly influenced by microcarrier concentrations and cell culture growth. Higher microcarrier concentrations led to higher average shear stress and Kolmogorov values. The cultured HEK-293 cells after seven days of growth also had higher average shear stress and Kolmogorov values than at the day of seeding, indicating an impact caused by the cells’ metabolism and biomass. Overall, the results indicated that assuming water-like properties underestimates shear stress and Kolmogorov length scales, especially at zones of lower shear rates due to the observed shear thinning behavior. Thus, careful monitoring of dynamic viscosity of cell cultures and proper control of mixing parameters are critical to deliver the desired mixing conditions for optimized cell growth especially during scale-up production operations.

目前已开发出计算流体动力学（CFD）模型来模拟细胞培养生物反应器，但由于数据严重不足，这些模型假定具有类似水的粘度特性。本研究描述了 HEK-293 细胞培养物的动态粘度，并评估了其对旋转瓶生物反应器中混合性能的影响。粘度测量结果表明，细胞培养基、含有微载体的培养基和细胞培养物在 1 到 100 s-1 的测量剪切速率范围内都表现出剪切变稀的行为。粘度还随着微载体浓度和细胞培养物的生长而增加。包含动态粘度数据的 CFD 模型显示，微载体浓度和细胞培养物生长对剪切应力和科尔莫哥罗夫长度曲线有显著影响。微载体浓度越高，平均剪切应力和 Kolmogorov 值越高。生长七天后的 HEK-293 细胞的平均剪切应力和 Kolmogorov 值也高于播种当天，这表明细胞的新陈代谢和生物量造成了影响。总之，研究结果表明，假设细胞具有类似水的特性会低估剪切应力和 Kolmogorov 长度尺度，尤其是在剪切速率较低的区域，这是因为观察到了剪切变稀行为。因此，仔细监测细胞培养物的动态粘度和适当控制混合参数对于提供理想的混合条件以优化细胞生长至关重要，尤其是在放大生产操作过程中。

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引用次数: 0

Integrating azo dye degradation and lipid accumulation by Candida tropicalis and Pichia kudriavzevii along with insights into underlying metabolomics for treatment of textile effluents 整合热带念珠菌和 Pichia kudriavzevii 降解偶氮染料和积累脂质的方法，以及对潜在代谢组学的深入了解，以处理纺织污水

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-09 DOI: 10.1016/j.bej.2024.109521

Sadik Dantroliya, Pooja Doshi, Ishan Raval, Chaitanya Joshi, Madhvi Joshi

Azo compounds, extensively utilized across various industries, contribute to the release of toxic effluents that are detrimental to both the environment and human health. Traditional methods for azo dye removal often result in harmful byproducts or concentrated sludge, complicating disposal efforts. This study explores the potential of two yeast strains, Candida tropicalis and Pichia kudriavzevii, to effectively decolorize azo dyes (TD4, TD5, and TD6) while simultaneously accumulating lipids. The cultures achieved 80–90 % decolorization of the selected dyes during incubation, with Pichia showing higher efficiency across multiple dyes compared to Candida. Lipid profiling identified valuable fatty acids, such as palmitic acid and oleic acid, with potential applications in biofuels and other industries. Total Organic Carbon (TOC) analysis revealed a reduction in TOC, indicating degradation and mineralization of the dyes by the yeasts. Metabolic profiling via LC-MS confirmed the degradation, showing the presence of intermediates such as azoles, azolines, isoquinolines, pyridines, and benzopyrans in dye-supplemented cultures. Additionally, pathways related to energy metabolism, amino acid metabolism, drug metabolism (cytochrome P450), degradation of aromatic compounds, and steroid biosynthesis were enriched in the dye-treated cultures. Lipid output in the presence of dyes ranged from 40 % to 90 %. The study thus demonstrates a proof of concept for economically viable lipid production combined with efficient dye removal, presenting a sustainable solution to environmental and industrial challenges.

各行各业广泛使用的偶氮化合物会释放出对环境和人类健康有害的有毒废水。传统的偶氮染料去除方法往往会产生有害的副产品或浓缩污泥，使处理工作变得更加复杂。本研究探讨了热带念珠菌和 Pichia kudriavzevii 这两种酵母菌株在积累脂质的同时有效脱色偶氮染料（TD4、TD5 和 TD6）的潜力。在培养过程中，这些培养物对所选染料的脱色率达到 80-90%，与念珠菌相比，Pichia 对多种染料的脱色效率更高。脂质分析确定了有价值的脂肪酸，如棕榈酸和油酸，它们在生物燃料和其他行业具有潜在的应用价值。总有机碳 (TOC) 分析显示 TOC 有所减少，表明酵母菌对染料进行了降解和矿化。通过液相色谱-质谱（LC-MS）进行的代谢分析证实了降解，显示在添加染料的培养物中存在唑类、偶氮啉类、异喹啉类、吡啶类和苯并吡喃类等中间产物。此外，与能量代谢、氨基酸代谢、药物代谢（细胞色素 P450）、芳香族化合物降解和类固醇生物合成有关的途径也在染料处理的培养物中得到了丰富。在有染料存在的情况下，脂质产量从 40% 到 90% 不等。因此，该研究证明了经济上可行的脂质生产与高效染料去除相结合的概念，为环境和工业挑战提供了一种可持续的解决方案。

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引用次数: 0

Combined optical measurement of dissolved oxygen tension (DOT), pH value, biomass and viscosity in shake flasks 在摇瓶中对溶解氧张力 (DOT)、pH 值、生物量和粘度进行组合光学测量

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-05 DOI: 10.1016/j.bej.2024.109515

Carl Dinter , David Vonester , David Flitsch , Moritz Mertens , Marc Tüschenbönner , Maximilian Hoffmann , Jochen Büchs , Jørgen Magnus

Shake flasks are widely spread in microbial process development. Characterization of the processes by manual offline sampling is time-consuming, highly laborious and a contamination risk. Online monitoring of key parameters would provide deeper insights, while saving time and effort. In this study, a device for optical online monitoring of dissolved oxygen tension (DOT), biomass, pH value and viscosity in shake flasks is presented. DOT measurement relies on fluorescent oxygen sensitive nanoparticles. The fluorescence intensity signal of the nanoparticles is used to trigger the DOT and scattered light measurements inside the rotating bulk liquid. The scattered light signal (610 – 630 nm) can be correlated to offline measured optical density OD₆₀₀, even at elevated viscosity. The pH value is monitored online by using pH sensor spots, fixed inside the shake flasks. The shift of the angle of the bulk liquid Θ-Θ₀ is correlated to the offline measured viscosity. Detection of the leading edge of the bulk liquid, necessary for viscosity measurement, can be performed either using the fluorescence intensity signal of the oxygen nanoparticles or the scattered light signal.

摇瓶广泛应用于微生物工艺开发。通过人工离线取样来描述工艺流程既费时又费力，还存在污染风险。对关键参数进行在线监测可提供更深入的见解，同时节省时间和精力。本研究介绍了一种用于光学在线监测摇瓶中溶解氧张力（DOT）、生物量、pH 值和粘度的装置。溶解氧张力测量依赖于荧光氧敏感纳米粒子。纳米颗粒的荧光强度信号用于触发旋转液体中的 DOT 和散射光测量。即使在粘度较高的情况下，散射光信号（610 - 630 nm）也可与离线测量的光密度 OD600 相关联。pH 值通过固定在摇瓶内的 pH 传感器点进行在线监测。散装液体 Θ-Θ0 角度的变化与离线测量的粘度相关。粘度测量所需的散装液体前缘检测可通过氧纳米粒子的荧光强度信号或散射光信号进行。

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引用次数: 0

Oxygen uptake rate analysis to evaluate the impact of hydrodynamic stress on the growth of the avian cell line DuckCelt®-T17 通过摄氧量分析评估流体动力压力对禽类细胞系 DuckCelt®-T17 生长的影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-05 DOI: 10.1016/j.bej.2024.109520

Valentine Tingaud , Philippe Lawton , Johan Peralez , Madiha Nadri-Wolf , Isabelle Pitault , Claudia Cogné , Elisabeth Errazuriz , Eyad Al Mouazen , Claire Bordes

Scale-up of bioprocesses involving animal cell culture is hampered by the sensitivity of the cells to hydrodynamic stress, either from agitation or bubble bursting. Here, the hydrodynamic stress experienced by a recent cell line, the DuckCelt®-T17 avian cells, previously used for viral vaccine production, is investigated in shake flasks and in a 3 L bioreactor. Cell stress was assessed by monitoring the dissolved oxygen in the culture medium, which depends on Oxygen Transfer Rate (OTR) and Oxygen Uptake Rate (OUR) during cultivation. Classical parameters such as the maximum growth rate (µ_max) and metabolite profiles were also determined. A dynamic model able to predict nutrient consumption, metabolic waste production, viable cell number and OUR was also developed and validated from the data measured in shake flasks. The experiments performed in the stirred tank bioreactor (STBR) show that OUR depended on both the cell growth phase and the stirring conditions. The oxygen consumption of the cells during the exponential growth phase (where there were no nutrient and O₂ limitations) was significantly altered at average and maximum shear rates above 70 and 840 s⁻¹, respectively, indicating highly shear-sensitive cells. OUR is a suitable tool to identify the hydrodynamic conditions for robust cell growth. The scale-up criteria to be favored for the DuckCelt®-T17 cell culture in STBRs would be the shear and/or the tip’s speed.

由于细胞对搅拌或气泡破裂产生的流体动力应力非常敏感，因此影响了涉及动物细胞培养的生物工艺的放大。在这里，我们研究了最近的一种细胞系--DuckCelt®-T17 禽细胞（以前曾用于病毒疫苗的生产）在摇瓶和 3 L 生物反应器中经历的流体动力应力。细胞压力通过监测培养基中的溶解氧来评估，溶解氧取决于培养过程中的氧转移率（OTR）和氧吸收率（OUR）。此外，还测定了最大生长速率（µmax）和代谢物概况等经典参数。此外，还开发了一个动态模型，可预测营养物质消耗、代谢废物产生、存活细胞数和 OUR，并根据摇瓶中测量的数据进行了验证。在搅拌罐生物反应器（STBR）中进行的实验表明，OUR 取决于细胞生长阶段和搅拌条件。当平均剪切速率超过 70 s-1 和最大剪切速率超过 840 s-1 时，细胞在指数生长阶段（没有营养和氧气限制）的耗氧量会发生显著变化，这表明细胞对剪切非常敏感。OUR 是确定细胞稳健生长的流体动力学条件的合适工具。在 STBR 中进行 DuckCelt®-T17 细胞培养的放大标准是剪切力和/或尖端速度。

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引用次数: 0

Streamlining process development and scale-up: Risk assessment to reduce workload in primary protein recovery 简化工艺开发和放大：风险评估以减少初级蛋白质回收的工作量

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-05 DOI: 10.1016/j.bej.2024.109513

Markus C. Berg , Yvonne Sorz , Rainer Hahn , Michael C. Martinetz , Cécile Brocard , Astrid Dürauer

Risk assessment is an integral aspect of the Quality-by-Design strategy to identify potential obstacles at every stage of biopharmaceutical production, from process development to tech transfer. We explored flow process chart, root cause analysis, and failure mode and effects analysis, to assess the scale-up of bacterial cell disruption and its influence on centrifugation and filtration steps. The Ishikawa diagram suggests that data on the impact of homogenizer valve design on product release, impurity profile, particle size distribution, viscosity, and dsDNA fragment size are missing which were collected experimentally for this study. Cell lysates from micro-, lab- and pilot scales cell disruption were analyzed for the above-mentioned parameters. Process parameters affecting these output parameters were identified on each individual scale. Cell disruption on the micro scale was performed in a bead mill. High pressure homogenization was used on lab- and pilot scales. Cell disintegration by bead milling delivers homogenates of product and impurity content comparable to those on bench scale but with 3-fold higher viscosity and significantly larger dsDNA fragments, 8.0 instead of 1.0 kbp, respectively. Miniaturized pressure flow curves identified dsDNA fragment sizes as critical for filter performance during clarification. Combining risk assessment, micro scale cell disintegration and bench scale pressure flow curves allows for selective and efficient process development, and scale up for primary recovery steps.

风险评估是 "质量源于设计 "战略不可或缺的一个方面，它可以识别从工艺开发到技术转让等生物制药生产各个阶段的潜在障碍。我们探索了流程图、根本原因分析和失效模式及影响分析，以评估细菌细胞破坏的放大及其对离心和过滤步骤的影响。石川图显示，本研究通过实验收集的有关均质器阀门设计对产品释放、杂质分布、粒度分布、粘度和 dsDNA 片段大小影响的数据缺失。对微量、实验室和中试规模细胞破碎的细胞裂解物进行了上述参数分析。确定了每个规模上影响这些输出参数的工艺参数。微量级细胞破碎是在珠磨机中进行的。高压均质法用于实验室和中试规模。通过珠磨机进行细胞破碎，得到的匀浆产品和杂质含量与工作台规模的匀浆产品和杂质含量相当，但粘度高出 3 倍，dsDNA 片段明显更大，分别为 8.0 kbp 而不是 1.0 kbp。微型压力流量曲线确定了 dsDNA 片段大小对澄清过程中的过滤性能至关重要。将风险评估、微尺度细胞解体和台架尺度压力流量曲线结合起来，可以进行选择性和高效的工艺开发，并扩大初级回收步骤的规模。

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引用次数: 0

A circular economy approach for utilization of tannery fleshing hydrolysate and kitchen wastes into organic fertilizer through enzymatic decomposition 通过酶分解将制革肉水解物和厨房废物利用为有机肥料的循环经济方法

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-02 DOI: 10.1016/j.bej.2024.109519

Md. Mazharul Islam , Fatema Tujjohra , Uttam K. Roy , Mohammed Mizanur Rahman

This study presents a sustainable method for converting tannery fleshing waste (FH) into organic fertilizer using enzymatic decomposition with crude protease. After extracting fat from the enzymatic hydrolysis, the enzyme-rich residue was mixed with dried kitchen waste (KW) and allowed to decompose for 45 days, producing nutrient-rich fertilizer. FT-IR spectroscopy confirmed the presence of important functional groups, including hydroxyl, aliphatic hydrocarbons, esters, and amide-I linkages. The organic fertilizers had higher nutrient content, with nitrogen (1.08-1.67%), phosphorus (0.78–0.98 %), potassium (0.1–0.76 %), and magnesium (239–259.5 ppm) which is higher than commercial fertilizers. FESEM-EDX analysis revealed a dense, porous structure with a high surface concentration of calcium, which enhances nutrient release in the soil. Dissolution tests showed that nutrients from the organic fertilizer were released gradually over 36 hours, whereas commercial NPK fertilizers released nutrients within 150 minutes in simulated soil-water conditions. Field trials with a Randomized Complete Block Design (RCBD) demonstrated improved growth in Malabar spinach, particularly with fertilizer sample S-5, which had an optimal flesh to kitchen waste (FH to KW) ratio of 1:10. Although S-5 had a lower nitrogen content (1.08 %), it contained higher levels of phosphorus (0.98 %), potassium (0.765 %), and magnesium (259.5 ppm), contributing to enhanced plant growth. The organic fertilizer resulted in a shoot length of 38.8 ± 2.0 cm, root length of 16.33 cm, 31 ± 3 leaves, and 95.12 % dry matter of the plant. Heavy metal analysis of the plant confirmed that levels of chromium (Cr), iron (Fe), nickel (Ni), cadmium (Cd), copper (Cu), and lead (Pb) were within WHO safety limits. Phytotoxicity tests of the fertilizer also showed no negative impact on Malabar spinach seed germination.

本研究提出了一种可持续的方法，利用粗蛋白酶的酶解作用将制革厂的制革废料（FH）转化为有机肥料。从酶水解中提取脂肪后，富含酶的残留物与干燥的厨余垃圾（KW）混合，并让其分解 45 天，从而产生营养丰富的肥料。傅立叶变换红外光谱证实了重要官能团的存在，包括羟基、脂肪烃、酯和酰胺-I 连接。有机肥的养分含量较高，氮（1.08%-1.67%）、磷（0.78%-0.98%）、钾（0.1%-0.76%）和镁（239-259.5 ppm）的含量均高于商业肥料。FESEM-EDX 分析表明，钙的表面浓度高，结构致密多孔，从而促进了土壤中养分的释放。溶解试验表明，有机肥的养分在 36 小时内逐渐释放，而商业 NPK 肥料在模拟土壤-水条件下 150 分钟内就能释放养分。采用随机完全区组设计（RCBD）进行的田间试验表明，马拉巴尔菠菜的生长情况有所改善，尤其是肥料样品 S-5，它的最佳肥肉与厨余（FH 与 KW）比例为 1:10。虽然 S-5 的氮含量较低（1.08%），但它含有较高水平的磷（0.98%）、钾（0.765%）和镁（259.5 ppm），有助于促进植物生长。施用有机肥后，植物的芽长为（38.8 ± 2.0）厘米，根长为 16.33 厘米，叶片为（31 ± 3）片，干物质含量为 95.12%。对植物的重金属分析表明，铬（Cr）、铁（Fe）、镍（Ni）、镉（Cd）、铜（Cu）和铅（Pb）的含量均在世界卫生组织的安全范围之内。肥料的植物毒性测试也表明，对马拉巴尔菠菜种子的发芽没有负面影响。

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引用次数: 0

Strategies for enhanced acetone-butanol-ethanol production by Clostridium beijerinckii Y10 from glucose and xylose and exploration of its physiological mechanisms 增强贝氏梭菌 Y10 利用葡萄糖和木糖生产丙酮-丁醇-乙醇的策略及其生理机制探索

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-02 DOI: 10.1016/j.bej.2024.109518

Yujie Jing, Mengying Liu, Shuaiying Peng, Huanhuan Ding, Saijin Wei, Qinghua Zhang, Hanguang Li

To enhance the production of butanol and its ratio to the total solvent, the impacts of exogenous amino acids supplementation on acetone-butanol-ethanol (ABE) fermentation were studied. With this effort, the highest butanol and total solvent production of 14.18 ± 0.24 g/L and 16.29 ± 0.31 g/L were achieved when 5.0 mg/L of phenylalanine was added at 18 h. Additionally, to explore the physiological reasons for the efficient synthesis of butanol under the optimal condition, the changes of key enzyme activities in the butanol synthesis pathway and the intracellular microenvironment were investigated. The findings show that phenylalanine addition could enhance crude enzyme activities of acetate kinase and butyrate kinase activities, strengthen NADH synthesis and expend more ATP to promote the growth of Clostridium beijerinckii Y10. This study suggests that the method of a tiny amount of amino acid addition was a simple and efficient approach to enhance bio-butanol production, thus providing a new strategy to improve the performance of other similar fermentation.

为了提高丁醇的产量及其在总溶剂中的比例，研究了补充外源氨基酸对丙酮-丁醇-乙醇（ABE）发酵的影响。结果表明，在 18 小时内添加 5.0 mg/L 的苯丙氨酸时，丁醇和总溶剂的产量最高，分别为 14.18 ± 0.24 g/L 和 16.29 ± 0.31 g/L。此外，为了探索丁醇在最佳条件下高效合成的生理原因，研究了丁醇合成途径中关键酶活性的变化和细胞内微环境的变化。研究结果表明，苯丙氨酸的添加可提高乙酸激酶和丁酸激酶的粗酶活性，加强 NADH 的合成，消耗更多的 ATP，从而促进贝氏梭菌 Y10 的生长。这项研究表明，添加微量氨基酸是提高生物丁醇产量的一种简单而有效的方法，从而为提高其他类似发酵的性能提供了一种新策略。

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引用次数: 0