Process Biochemistry最新文献_第4页

A review of genetic engineering techniques for CTG(Ser1) and CTG(Ala) D-xylose-metabolizing yeasts employed for second-generation bioethanol production 用于第二代生物乙醇生产的 CTG（Ser1）和 CTG（Ala）D-木糖代谢酵母的基因工程技术综述

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-30 DOI: 10.1016/j.procbio.2024.09.028

Ana Paula Wives , Isabelli Seiler de Medeiros Mendes , Sofia Turatti dos Santos , Diego Bonatto

D-xylose is the second most abundant monosaccharide found in lignocellulose and is of biotechnological importance for producing second-generation ethanol and other high-value chemical compounds. D-xylose conversion to ethanol is promoted by microbial fermentation, mainly by bacteria, yeasts, or filamentous fungi. Among yeasts, species belonging to the CTG(Ser1) or CTG(Ala) clade display a remarkable ability to ferment D-xylose to ethanol and other compounds; however, these yeasts are not employed on an industrial scale given their poor fermentative performance compared to that of conventional yeasts, such as Saccharomyces cerevisiae, and because of the lack of a molecular toolbox for the development of new strains tailored to fermentation stress tolerance and performance. Thus, the purpose of this review was to evaluate the major genetic engineering tools (e.g., transformation markers and techniques, vectors, regulatory sequences, and gene editing techniques) available for the most studied yeasts of the CTG(Ser1) clade, such as Scheffersomyces, Spathaspora, Candida, and Yamadazyma species, and the CTG(Ala) clade, representative Pachysolen tannophilus. Furthermore, we systematized state-of-the-art molecular developments and perspectives to design D-xylose-fermenting yeast strains.

D- 木糖是木质纤维素中含量第二高的单糖，对生产第二代乙醇和其他高价值化合物具有重要的生物技术意义。D-木糖转化为乙醇的过程主要由细菌、酵母菌或丝状真菌等微生物发酵促进。在酵母菌中，属于 CTG（Ser1）或 CTG（Ala）支系的菌种具有将 D-木糖发酵成乙醇和其他化合物的卓越能力；然而，与传统酵母菌（如酿酒酵母）相比，这些酵母菌的发酵性能较差，而且缺乏针对发酵胁迫耐受性和性能开发新菌株的分子工具箱，因此没有在工业规模上使用这些酵母菌。因此，本综述的目的是评估 CTG（Ser1）支系（如 Scheffersomyces、Spathaspora、Candida 和 Yamadazyma 种）和 CTG（Ala）支系（具有代表性的 Pachysolen tannophilus）中研究最多的酵母菌可用的主要基因工程工具（如转化标记和技术、载体、调控序列和基因编辑技术）。此外，我们还系统地介绍了设计D-木糖发酵酵母菌株的最新分子发展和前景。

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引用次数: 0

Enhancement of abscisic acid biosynthesis in Saccharomyces cerevisiae via multidimensional engineering 通过多维工程提高赤霉酸在酿酒酵母中的生物合成能力

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-30 DOI: 10.1016/j.procbio.2024.09.029

Xiaofei Song , Jianze Zhang , Xikai Wang , Haonan Yu , Nuo Xu , Longyu Cao , Xiuwen Zhong , Puhong Yi , Jie Sun , Kun Wang , Chao Feng , Weixia Wang , Tingheng Zhu

Abscisic acid (ABA), a type of sesquiterpenoid plant hormone, has high application value in agriculture, nutrition and medicine. Herein, we constructed an efficient ABA-producing yeast cell factory by combining multidimensional engineering strategies. Starting from a suitable strain, YS010 was selected from 11 varieties of S. cerevisiae strains by evaluating ergosterol content and growth ability, then the biosynthetic pathway of ABA derived from Botris cinerea was constructed, resulting in 1.93 mg L⁻¹ ABA. Next, the metabolic flux of the mevalonic acid (MVA) pathway was increased to enhance the synthesis of the precursor farnesyl pyrophosphate (FPP). To further increase the FPP competitiveness of the ABA synthesis pathway, we attempted to enhance the catalytic performance of BcABA3 through enzyme engineering, and the ABA yield of the mutant strain YS036-ABAP^A206D reached 2.64 mg L⁻¹ in SC-ura medium. In addition, we developed a multi-copy integration strategy, TPI1-delta driven CRISPR-Cas9 (TDI-CRISPR) integration system, to realize the high copy and stable expression of bcaba1, bcaba2 and bcaba3, which enabled the titer of ABA to reach 17.47 mg L⁻¹. Finally, by optimising the fermentation medium, the ABA titer reached 30.30 mg L⁻¹, which was the highest level ever reported for de novo ABA biosynthesis in S. cerevisiae.

脱落酸（ABA）是一种倍半萜类植物激素，在农业、营养学和医学方面具有很高的应用价值。在此，我们结合多维工程策略，构建了一个高效的ABA生产酵母细胞工厂。首先，通过评估麦角甾醇含量和生长能力，从 11 个 S. cerevisiae 菌株品种中筛选出一株合适的菌株 YS010，然后构建了从 Botris cinerea 提取 ABA 的生物合成途径，得到 1.93 mg L-1 ABA。接着，增加了甲羟戊酸（MVA）途径的代谢通量，以提高前体焦磷酸法尼基（FPP）的合成。为了进一步提高 FPP 在 ABA 合成途径中的竞争力，我们尝试通过酶工程提高 BcABA3 的催化性能，结果突变株 YS036-ABAPA206D 在 SC-ura 培养基中的 ABA 产量达到了 2.64 mg L-1。此外，我们还开发了一种多拷贝整合策略--TPI1-delta驱动的CRISPR-Cas9（TDI-CRISPR）整合系统，实现了bcaba1、bcaba2和bcaba3的高拷贝和稳定表达，使ABA的滴度达到17.47 mg L-1。最后，通过优化发酵培养基，ABA 的滴度达到了 30.30 mg L-1，这是迄今为止报道的 S. cerevisiae 从头合成 ABA 的最高水平。

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引用次数: 0

Effects of suspended titanium dioxide (TiO2) nanoparticles on cake layer formation in submerged anaerobic membrane bioreactor (AnMBR) for landfill leachate treatment (LFL) 悬浮二氧化钛（TiO2）纳米颗粒对用于垃圾渗滤液处理的浸没式厌氧膜生物反应器（AnMBR）中滤饼层形成的影响

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-27 DOI: 10.1016/j.procbio.2024.09.022

Serdar Göçer , Binnaz Zeynep Zaimoğlu , Kevser Cırık

In recent years, TiO₂ NPs have attracted great attention among the semiconductors because of stability, commercial availability, and ease of preparation. For this reason, NPs are widely used in wastewater treatment and membrane bioreactor (MBRs) In this study, the effect of titanium dioxide (TiO2) nanoparticle material was investigated on both the landfill leachate (LFL) treatment and membrane fouling performance. The system performance was evaluated for under varying TiO₂ concentrations (50–300 mg/L TiO₂), constant HRT (24 h), and constant backwashing (5 min)-relaxing (0.5 min) in AnMBR. The optimum conditions were determined as 300 mg/L TiO₂ and the corresponding to COD, Color, TOC and TN removal efficiencies were observed as 55 %, 23 %, 22 %, 30 %, respectively. The best membrane performance was observed at 300 mg/L TiO₂ corresponding to membrane fouling rate as 0.01 mbar/min. TiO₂ addition significantly mitigated membrane fouling (75 % decrease) for AnMBR. Bacteroidetes, Firmicutes and Proteobacteria have been observed to be the dominant species in LFL and MBRs. The bacterial species responsible for membrane fouling were determined as Alphaproteobacteria, Sphingobacteria and Flavobacteria. The addition of TiO₂ was determined membrane fouling decreased in AnMBR. As a result of TiO₂ NPs were observed to thin the cake layer and postpone membrane fouling and filtration.

近年来，二氧化钛（TiO2）纳米粒子因其稳定性、商业可用性和易制备性而在半导体中备受关注。因此，纳米粒子被广泛应用于废水处理和膜生物反应器（MBRs）中。在 AnMBR 中，评估了不同 TiO2 浓度（50-300 mg/L TiO2）、恒定 HRT（24 小时）和恒定反冲洗（5 分钟）-松弛（0.5 分钟）条件下的系统性能。最佳条件确定为 300 mg/L TiO2，相应的 COD、色度、TOC 和 TN 去除率分别为 55 %、23 %、22 % 和 30 %。在 300 毫克/升 TiO2 的条件下，膜性能最佳，相应的膜堵塞率为 0.01 毫巴/分钟。添加 TiO2 能明显减轻 AnMBR 的膜堵塞（减少 75%）。据观察，类杆菌、固着菌和变形菌是低温低氧反应器和膜生物反应器中的优势菌种。造成膜堵塞的细菌种类被确定为兼性蛋白杆菌、鞘氨醇杆菌和黄杆菌。添加 TiO2 后，AnMBR 中的膜堵塞现象有所减少。结果表明，TiO2 NPs 可使滤饼层变薄，推迟膜堵塞和过滤。

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引用次数: 0

Preparation and characterization of novel antioxidant peptides from protein hydrolysate of Ophiocordyceps gracilis 从蛇床子草蛋白水解物中制备新型抗氧化肽并确定其特性

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-26 DOI: 10.1016/j.procbio.2024.09.026

Yidan Zhang, Hui Lian, Linhui Yang, Lingling Tong, Yang Wu, Suxing Jin, Dongsheng Guo

Ophiocordyceps gracilis is a type of fungus commonly utilized in both health food and Chinese medicine. Herein, we prepared antioxidant hydrolysate from O. gracilis, and isolated and purified the prepared hydrolysate to obtain O. gracilis peptides-4–3–1 (OGP-4–3–1) grade with the highest scavenging activity on DPPH, ABTS and hydroxyl radicals. Three new peptides with high antioxidant capacity were identified as LFNHF, PLDRHPF and YPFLRPL, respectively. The three peptides can reduce the proportion of senescent cells conveying positive expression of senescence-associated-β-galactosidase (SA-β-Gal) and increase survival pace of H₂O₂-induced human hepatocellular carcinomas (HepG2 cells), and the protective effects on cellular against senescence might be related to their increase in the activities of antioxidant enzymes, as well as the decrease in the concentration of malondialdehyde (MDA) and reactive oxygen species (ROS). The molecular docking results indicated that they could form durable hydrophobic and hydrogen bonds with critical amino acid residues of the Kelch-like ECH-associated protein 1 (Keap1), which could potentially modulate pathway of nuclear factor erythroid 2-related factor 2 (Nrf2) in conjunction with Keap1. In conclusion, the current research offers a theoretical foundation for the further development of the anti-aging effect of OGP, which has a broad application prospect in the fields of functional food and cosmetics.

麦角菌是一种常用于保健食品和中药的真菌。本文制备了蝙蝠蛾肽的抗氧化水解物，并对其进行分离纯化，得到了对DPPH、ABTS和羟基自由基具有最高清除活性的蝙蝠蛾肽-4-3-1（OGP-4-3-1）。经鉴定，三种新的高抗氧化能力多肽分别为LFNHF、PLDRHPF和YPFLRPL。这三种肽能降低衰老相关-β-半乳糖苷酶（SA-β-Gal）阳性表达的衰老细胞比例，提高H2O2诱导的人肝癌（HepG2细胞）的存活率，其对细胞抗衰老的保护作用可能与它们提高了抗氧化酶的活性、降低了丙二醛（MDA）和活性氧（ROS）的浓度有关。分子对接结果表明，它们能与 Kelch-like ECH-associated protein 1（Keap1）的关键氨基酸残基形成持久的疏水键和氢键，从而有可能与 Keap1 共同调节核因子红细胞 2 相关因子 2（Nrf2）的通路。总之，目前的研究为进一步开发 OGP 的抗衰老作用提供了理论基础，在功能食品和化妆品领域具有广阔的应用前景。

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引用次数: 0

Understanding in-vivo refolding of antibody fragments (Fab): Biosimilar Ranibizumab a case study 了解抗体片段（Fab）的体内再折叠：生物仿制药拉尼珠单抗案例研究

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-26 DOI: 10.1016/j.procbio.2024.09.027

Aatir A. Tungekar , Padmaja Fulewar , Rupali Kumthekar , Rahul Bhambure

In-vitro protein refolding is a major bottleneck step in the large-scale manufacturing of antibody fragments expressed in a microbial host system. The formation of an inter-chain disulfide bond is a key rate-limiting step during in-vitro refolding of antibody fragment therapeutics. In this investigation, we report the use of two redox mutant strains of E. coli viz., SHuffle® T7 (DE3) and SHuffle® T7 Express (DE3) possessing an oxidizing cytoplasm for the soluble expression of a refolded biosimilar antibody fragment. The effect of various critical process parameters on antibody fragment refolding yield was studied using a Design of Experiment (DoE) approach. The optimized upstream processing led to the expression of 167 ± 2.53 mg/L and 95 ± 1.44 mg/L of soluble refolded biosimilar Ranibizumab using the SHuffle® T7 (DE3) and SHuffle® T7 Express (DE3) strains, respectively, at the bioreactor scale. Physicochemical characteristics of the in-vivo refolded antibody fragment were studied and compared with an innovator molecule using various orthogonal analytical methods. A biological activity study of in-vivo refolded Ranibizumab using HUVEC cell-based bioassay proved that cell proliferation inhibition is comparable to the innovator Ranibizumab. The proposed strategy offers a time and cost-effective manufacturing platform for antibody fragments.

体外蛋白质重折叠是大规模生产在微生物宿主系统中表达的抗体片段的主要瓶颈步骤。链间二硫键的形成是抗体片段治疗体外重折叠过程中的关键限速步骤。在这项研究中，我们报告了使用两种氧化还原突变的大肠杆菌菌株，即 SHuffle® T7 (DE3) 和 SHuffle® T7 Express (DE3)，它们具有氧化细胞质，用于可溶性表达再折叠的生物类似抗体片段。实验设计（DoE）方法研究了各种关键工艺参数对抗体片段重折叠产量的影响。通过优化上游处理，使用 SHuffle® T7 (DE3) 和 SHuffle® T7 Express (DE3) 菌株在生物反应器规模上分别表达了 167 ± 2.53 mg/L 和 95 ± 1.44 mg/L 的可溶性再折叠生物仿制药雷珠单抗。使用各种正交分析方法研究了体内重折叠抗体片段的理化特性，并与创新分子进行了比较。使用基于 HUVEC 细胞的生物测定法对体内重折叠的雷珠单抗进行的生物活性研究证明，其细胞增殖抑制作用与创新药雷珠单抗相当。所提出的策略为抗体片段的生产提供了一个省时、经济的平台。

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引用次数: 0

Interaction between the electrochemical properties of powdered activated carbon and the biochemical processes within bacteria in Azo dye biodecolorization: An explanatory mechanism 偶氮染料生物脱色过程中粉末活性炭的电化学特性与细菌内部生化过程之间的相互作用：一种解释机制

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-26 DOI: 10.1016/j.procbio.2024.09.025

Pedram Hassanvand , Masoud Rajabi , Tayebe Bagheri Lotfabad , Soheila Yaghmaei

Drawing on prior reports highlighting the redox mediator properties of powdered activated carbon (PAC), this study was designed to evaluate these properties to enhance the decolorization of azo dye by Klebsiella quasipneumoniae GT7. It was found that the presence of 0.5 % PAC in the medium increased the biodecolorization rate early in incubation. Chemical analysis revealed that dye conversion into aromatic amines occurred in microbial systems both with and without PAC. However, at initial dye concentrations (

C_{id}

) of 2 mM or higher, some dye remained on the PAC surface and in the medium. In contrast, the PAC-free system achieved nearly 100 % biodecolorization at all initial dye concentrations. The negative impact of PAC on decolorization efficiency in microbial systems with high initial dye concentrations cannot be solely explained by its redox mediator function. This study used the amphoteric-Donnan model for PAC's electrical double layer (EDL) and Mitchell's chemiosmotic model for bacterial proton motive force (PMF) to explore this. It found that charge storage in PAC's EDL regulates electron transfer fluxes, and proton species enhance the proton motive force across the bacterial membrane. These observations improve the understanding of PAC's role in microbial decolorization and its potential future applications.

先前的报告强调了粉末活性碳（PAC）的氧化还原介质特性，本研究旨在评估这些特性，以提高准肺炎克雷伯氏菌 GT7 对偶氮染料的脱色能力。研究发现，在培养基中加入 0.5 % 的 PAC 可提高培养初期的生物脱色率。化学分析显示，在有 PAC 和没有 PAC 的微生物系统中，染料都会转化为芳香胺。然而，当初始染料浓度（Cid）为 2 mM 或更高时，一些染料会残留在 PAC 表面和培养基中。相比之下，不含 PAC 的系统在所有初始染料浓度下都几乎实现了 100% 的生物脱色。在初始染料浓度较高的微生物体系中，PAC 对脱色效率的负面影响不能完全由其氧化还原介质功能来解释。本研究利用 PAC 的双电层（EDL）两性-多南模型和 Mitchell 的细菌质子动力（PMF）化学渗透模型对此进行了探讨。研究发现，PAC 双电层中的电荷储存调节着电子传递通量，质子种类增强了细菌膜上的质子动力。这些观察结果加深了人们对 PAC 在微生物脱色过程中的作用及其未来潜在应用的理解。

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引用次数: 0

Antioxidant and anticancer activities on HT-29 colon cancer cells of protein isolate extracted from Cordyceps militaris fruiting body using diverse isolation methods 采用不同分离方法从冬虫夏草子实体中提取的分离蛋白对 HT-29 结肠癌细胞的抗氧化和抗癌活性

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-24 DOI: 10.1016/j.procbio.2024.09.024

Mongkol Thirabunyanon , Natthapong Mungmueang , Wichittra Daengprok , Chitraporn Ngampeerapong , Supatra Karnjanapratum , Soottawat Benjakul , Sirin Panyakom , Theeraphol Senphan , Chodsana Sriket

The present research examines different techniques for isolating proteins from Cordyceps militaris fruiting body powder (COR), namely ammonium sulfate precipitation (ASP), organic solvent precipitation (OSP), and isoelectric precipitation (IEP). ASP yielded the highest protein content (78.90 %), effectively concentrating proteins, while OSP and IEP also produced substantial yields (57.41 % and 69.06 %, respectively). Color analysis revealed differences among isolates, with ASP resulting in a darker hue. ASP and IEP showed higher red and yellow components, respectively. Essential amino acid content was significantly higher in ASP isolate compared to COR, indicating effective amino acid concentration. Thermal analysis revealed different decomposition temperatures and enthalpy values among isolates. OSP displayed the highest phenolic content, while IEP exhibited the highest flavonoid content. Antioxidant assays showed ASP to have the highest DPPH-RSA and ABTS-RSA, while IEP showed the highest FRAP value. ASP demonstrated anticancer activity against HT-29 colon cancer cells inducing cell death apoptosis and inhibiting cell migration, with an IC₅₀ of 360.63 µg/mL, comparable to Paclitaxel. These findings suggest that Cordyceps militaris protein isolate, particularly from ASP, has significant antioxidant and anticancer potential, supporting its potential as a natural alternative to synthetic drugs with fewer side effects.

本研究探讨了从冬虫夏草子实体粉（COR）中分离蛋白质的不同技术，即硫酸铵沉淀法（ASP）、有机溶剂沉淀法（OSP）和等电沉淀法（IEP）。硫酸铵沉淀的蛋白质含量最高（78.90%），有效浓缩了蛋白质；有机溶剂沉淀和等电沉淀的蛋白质含量也很可观（分别为 57.41% 和 69.06%）。颜色分析显示了不同分离物之间的差异，ASP 的颜色较深。ASP 和 IEP 的红色和黄色成分分别较高。与 COR 相比，ASP 分离物的必需氨基酸含量明显较高，表明氨基酸浓度较高。热分析表明，不同分离物的分解温度和焓值不同。OSP 的酚含量最高，而 IEP 的类黄酮含量最高。抗氧化测定显示，ASP 的 DPPH-RSA 和 ABTS-RSA 值最高，而 IEP 的 FRAP 值最高。ASP 对 HT-29 结肠癌细胞具有抗癌活性，可诱导细胞凋亡并抑制细胞迁移，其 IC50 值为 360.63 µg/mL，与紫杉醇相当。这些研究结果表明，冬虫夏草蛋白分离物（尤其是 ASP）具有显著的抗氧化和抗癌潜力，支持其作为副作用较少的合成药物天然替代品的潜力。

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引用次数: 0

Pentagalloyl glucose enhanced the stress resistance to delay aging process in Caenorhabditis elegans 五聚酰基葡萄糖增强草履虫的抗应激能力以延缓衰老过程

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-24 DOI: 10.1016/j.procbio.2024.09.021

Jingrui Luo , Xiaoying Zhang , Wei Li , Yaqi Lan , Fangwen Li , Jie Xiao , Yong Cao , Guo Liu , Yunjiao Chen

Aging is a complex biological process characterized by gradual and irreversible functional deterioration, strongly associated with oxidative stress. Pentagalloyl glucose (PGG) has attracted increasing attention due to its potent antioxidant and anti-stress properties. This study investigated the potential of PGG to mitigate the aging process under stress in RAW 264.7 cells and Caenorhabditis elegans models. The expression of vital genes associated with stress was also measured to explain the action mechanism of PGG in C. elegans. The findings showed that PGG supplementation not only significantly enhanced the stress tolerance of RAW 264.7 cells, but also prolonged lifespan and reduced the ROS and lipofuscin accumulation in C. elegans induced by stress. Meanwhile, the improvement effect of PGG on delaying aging development was also manifested in the protection of mitochondrial function and neuronal integrity. Moreover, daf-16 nuclear translocation and sod-3 expression were significantly enhanced by PGG to delay the aging process. Mechanistically, PGG might alleviate aging by improving daf-16, sod-3, ctl-1, and gst-4 levels in the DAF-16/FOXO pathway and upregulating skn-1 and gst-4 expression in the SKN-1/Nrf2 pathway. Our study provided novel insights into the role of PGG in combating stress-induced aging.

衰老是一个复杂的生物过程，其特点是功能逐渐退化且不可逆转，与氧化应激密切相关。五聚酰基葡萄糖（Pentagalloyl glucose，PGG）因其强大的抗氧化和抗应激特性而受到越来越多的关注。本研究调查了 Pentagalloyl glucose 在 RAW 264.7 细胞和秀丽隐杆线虫模型中缓解压力下衰老过程的潜力。研究还测量了与应激相关的重要基因的表达，以解释 PGG 在秀丽隐杆线虫中的作用机制。研究结果表明，补充 PGG 不仅能显著增强 RAW 264.7 细胞的应激耐受性，还能延长秀丽隐杆线虫的寿命，减少应激诱导的 ROS 和脂褐素积累。同时，PGG 对延缓衰老发展的改善作用还体现在保护线粒体功能和神经元完整性方面。此外，PGG还能显著增强daf-16的核转位和sod-3的表达，从而延缓衰老过程。从机理上讲，PGG可能通过改善DAF-16/FOXO通路中的daf-16、sod-3、ctl-1和gst-4水平以及上调SKN-1/Nrf2通路中的skn-1和gst-4表达来缓解衰老。我们的研究为了解 PGG 在抗应激诱导的衰老中的作用提供了新的见解。

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引用次数: 0

Insight into microorganisms and flavor substances in traditional Chinese fermented food starter: Daqu 洞察中国传统发酵食品开胃菜中的微生物和风味物质：大曲

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-24 DOI: 10.1016/j.procbio.2024.09.015

Akhtiar Ali , Yanfang Wu , Weiwei Li , Zhongfu Duan , Ru Zhang , Jianing Liu , Prasanna J. Patil , Haroon Shah , Xiuting Li

Daqu, a crucial fermentation starter in the production of various Chinese fermented foods, plays a pivotal role in shaping complex enzyme profiles and diverse flavour precursors. This review aims to elucidate the microbial communities within Daqu, focusing on their functionalities in the context of flavour development. We delve into the detection methods of microorganisms and flavour substances in Daqu, employing advanced technologies including high-performance liquid chromatography, gas chromatography, pseudo-targeted metabolomics, and headspace solid-phase microextraction-gas chromatography-mass spectrometry. This review explores high throughput sequencing technologies, including pyrosequencing, clonal library sequencing, metaproteomic, and metagenomics, to gain a comprehensive understanding of the intricate microbial dynamics. Additionally, we discuss the metabolic pathways involved in flavour substance production within Daqu. By synthesizing information on Daqu types, microorganisms present, detection methodologies, and flavour substance metabolic pathways, this review contributes to a deeper comprehension of the intricate processes underpinning the Flavors of Chinese fermented foods.

大曲是生产各种中国发酵食品的重要发酵起始菌，在形成复杂的酶谱和多种风味前体方面发挥着关键作用。本综述旨在阐明大曲中的微生物群落，重点关注它们在风味形成过程中的功能。我们深入探讨了大曲中微生物和风味物质的检测方法，采用的先进技术包括高效液相色谱法、气相色谱法、伪靶向代谢组学以及顶空固相微萃取-气相色谱-质谱联用技术。本综述探讨了高通量测序技术，包括热测序、克隆文库测序、元蛋白组学和元基因组学，以全面了解错综复杂的微生物动态。此外，我们还讨论了大曲中香味物质生产所涉及的代谢途径。通过综合大曲类型、存在的微生物、检测方法和风味物质代谢途径等方面的信息，本综述有助于加深对中国发酵食品风味复杂过程的理解。

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引用次数: 0

Understanding Bacillus response to salt stress: Growth inhibition, enhanced EPS secretion, and molecular adaptation mechanisms 了解芽孢杆菌对盐胁迫的反应：生长抑制、EPS 分泌增强和分子适应机制

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2024-09-24 DOI: 10.1016/j.procbio.2024.09.023

Zheng-yan Yin , Yi-cheng Yuan , Rui Zhang , Jun-ting Gan , Lei Yu , Xu-hai Qiu , Rong-ping Chen , Quan Wang

This study investigates the secretion pattern of extracellular polymeric substances (EPS) by Bacillus sp. under varying salt concentrations and elucidates the molecular mechanisms governing EPS synthesis and secretion. Salt stress inhibited cell proliferation, while optimal salt stimulation promoted EPS secretion, resulting in increased viscosity of the culture medium and the formation of bacterial clusters. Fourier infrared spectrum analysis revealed functional groups such as C-O-C and N-H within the EPS. Soluble-EPS (S-EPS) contained sulfur and phosphorus groups associated with heavy metal ions adsorption. The study also identified a novel polysaccharide formed through bonding EPS (B-EPS). High salt concentrations correlated with elevated levels of tryptophan protein and its derivatives, increased tyrosine polysaccharide derivatives, and decreased aromatic polysaccharides. B-EPS exhibited higher levels of aromatic polysaccharides, with Na⁺ promoting detachment of B-EPS from the cell surface. Transcriptome sequencing (RNA-seq) analysis under salt stress revealed significant expression of spore kinase (KinD) and response regulatory protein Spo0A in the phosphoric acid relay system. Key transcriptional regulatory factors, including OmpR and exopolysaccharide biosynthesis, were closely associated with EPS synthesis and secretion. This study establishes a theoretical foundation for the industrial production and practical application of EPS by elucidating the molecular mechanisms underlying Bacillus' response to salt stress.

本研究探讨了芽孢杆菌在不同盐浓度下分泌胞外聚合物物质（EPS）的模式，并阐明了EPS合成和分泌的分子机制。盐胁迫抑制了细胞增殖，而最佳盐刺激则促进了 EPS 的分泌，导致培养基粘度增加并形成细菌簇。傅立叶红外光谱分析揭示了 EPS 中的 C-O-C 和 N-H 等官能团。可溶性 EPS（S-EPS）含有与重金属离子吸附有关的硫和磷基团。研究还发现了一种通过粘合 EPS（B-EPS）形成的新型多糖。高浓度盐与色氨酸蛋白及其衍生物水平升高、酪氨酸多糖衍生物增加和芳香族多糖减少相关。B-EPS的芳香族多糖含量较高，Na+可促进B-EPS从细胞表面脱离。盐胁迫下的转录组测序（RNA-seq）分析显示，磷酸中继系统中的孢子激酶（KinD）和响应调控蛋白 Spo0A 有显著表达。包括 OmpR 和外多糖生物合成在内的关键转录调控因子与 EPS 的合成和分泌密切相关。本研究通过阐明芽孢杆菌对盐胁迫响应的分子机制，为 EPS 的工业化生产和实际应用奠定了理论基础。

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