Process Biochemistry最新文献_第6页

Sustainable 3D-printed immobilization system with soybean meal coating for high-yield Lactococcus lactis bioprocessing 用于高产乳酸乳球菌生物加工的可持续3d打印豆粕包衣固定系统

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-21 DOI: 10.1016/j.procbio.2025.10.009

Yi-Cheng Huang , Darin Khumsupan , Shin-Ping Lin , Kaemwich Jantama , Shella Permatasari Santoso , Chang-Wei Hsieh , Hui-Wen Lin , Kuan-Chen Cheng

This study utilized 3D-printed carriers coated with soybean meal, an agricultural by-product, to optimize the immobilization of Lactococcus lactis subsp. lactis, enhancing bacteriocin production and efficiency. Among the six experimental groups, the soybean-only (S) group achieved the highest yield of 1275.36 ± 17.49 IU/mL within two hours, which was 246 % higher than the non-coated group and significantly greater than the control (C) group (no 3D-printed carriers) (non-detect). After eight hours, the S group’s production rate was 2191.10 ± 21.24 IU·mL⁻¹ ·h⁻¹ , representing a 42.68 ± 2.20 % increase compared to the C group and 25.41 ± 1.80 % compared to the NC group. Under repeated batch fermentation, the S-only group achieved the highest total yield of 170,606.61 ± 3896.23 IU/mL over 10 cycles of shake flask fermentation, which was 20.42 % higher than static cultivation. Furthermore, after nine cycles, 72.2 % of activity was retained. The immobilized carrier also demonstrated improved hydrophilicity, with the water contact angle decreasing from 92.37° to 25.03°. SEM and FTIR analyses further confirmed the successful immobilization. Moreover, real-time RT-PCR analysis indicated that the increase in bacteriocin production was associated with the upregulation of 11 nisin-related genes, with nisA and nisB increasing by 4.74- and 4.86-fold, respectively. This study presents a novel approach for producing nisin by immobilizing Lactococcus lactis on 3D-printed structures coated with agricultural waste, demonstrating that immobilization can influence gene regulation. This technique offers a novel approach to valorizing agricultural waste while promoting environmentally responsible and economically viable industrial practices.

本研究利用3d打印载体包被豆粕（一种农业副产品），优化了乳酸乳球菌亚种的固定化。乳酸，提高细菌素的产量和效率。6个实验组中，仅大豆(S)组在2小时内的产率最高，为1275.36 ± 17.49 IU/mL，比未包被组高246 %，显著高于对照组(C)组（无3d打印载体）（未检测）。八小时后,S集团的产量是2191.10 ± 21.24 IU·mL⁻¹ ·h⁻¹ ,代表 42.68±2.20  %增加 相比C组和25.41±1.80  %相比数控组。在重复分批发酵条件下，S-only组在摇瓶发酵10个周期后，总产率最高，为170,606.61 ± 3896.23 IU/mL，比静态培养提高20.42 %。此外，经过9个循环，72.2 %的活性保留。固定化载体的亲水性也得到了改善，水接触角从92.37°降低到25.03°。SEM和FTIR分析进一步证实了固定化的成功。此外，实时RT-PCR分析表明，细菌素产量的增加与11个nisin相关基因的上调有关，其中nisA和nisB分别增加了4.74倍和4.86倍。本研究提出了一种新的方法，通过将乳酸乳球菌固定在涂有农业废弃物的3d打印结构上产生乳酸链球菌素，表明固定可以影响基因调控。这项技术提供了一种新的方法，在促进对环境负责任和经济上可行的工业实践的同时，对农业废物进行估价。

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引用次数: 0

Sustainable extraction of marine collagen and hydrolysates: Recent advances and challenges towards commercialization 海洋胶原蛋白和水解物的可持续提取：最近的进展和商业化的挑战

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-20 DOI: 10.1016/j.procbio.2025.10.005

Farhana Islam , Tanjina Tarannum , Mahdia Mahmud , Fahim Rahman , Nafisa Islam

Functional bioactive properties of collagen and its hydrolysates have boosted widespread applications of collagenous products extracted from natural sources. Growing interest in marine collagen has emerged due to its unique biochemical properties, compatibility with religious and cultural restrictions, and concerns over diseases linked to mammalian collagen. Furthermore, high availability of discarded marine by-products has facilitated various extraction techniques leading to conversion of low-cost by-products (e.g., fish skin, bones, scales, etc.) into high-value collagenous derivatives. Many innovative routes are being explored at different stages of extraction to enable sustainable production of bioactive collagens from marine sources, but most of these approaches remain at an early stage of development. This review summarizes conventional, enzymatic, physical-assisted, and green solvent-based methods for marine collagen and hydrolysate production, briefly discussing marine collagen market, challenges associated with commercializing marine collagen-based products, and highlighting opportunities to provide a clearer perspective. Enzyme-assisted methods, when coupled with physical- or solvent-assisted methods, balance yield, bioactivity, and sustainability, though large-scale validation is limited. Although this field has progressed significantly, widescale unified comparisons of bioactivity and yield are lacking. Future research should integrate techno-economic and life cycle analysis with pilot-scale studies, paving the way for industrial adoption of sustainable marine collagen.

胶原蛋白及其水解物的功能性生物活性特性促进了从天然来源提取的胶原产品的广泛应用。由于其独特的生化特性、与宗教和文化限制的相容性以及对与哺乳动物胶原蛋白有关的疾病的担忧，人们对海洋胶原蛋白的兴趣日益浓厚。此外，废弃海洋副产品的高可得性促进了各种提取技术，从而将低成本副产品（例如鱼皮、鱼骨、鱼鳞等）转化为高价值的胶原衍生物。在不同的提取阶段，正在探索许多创新路线，以便从海洋来源可持续地生产生物活性胶原蛋白，但这些方法中的大多数仍处于早期发展阶段。本文综述了传统的、酶促的、物理辅助的和绿色溶剂基的海洋胶原蛋白和水解产物的生产方法，简要讨论了海洋胶原蛋白的市场，与海洋胶原蛋白产品商业化相关的挑战，并强调了提供更清晰观点的机会。酶辅助方法，当与物理或溶剂辅助方法相结合时，平衡产量，生物活性和可持续性，尽管大规模验证是有限的。尽管这一领域取得了重大进展，但缺乏对生物活性和产量进行广泛统一的比较。未来的研究应该将技术经济和生命周期分析与中试研究结合起来，为可持续海洋胶原蛋白的工业应用铺平道路。

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引用次数: 0

Whole-cell biotechnological applications with thermoacidophilic Crenarchaeota: Opportunities and challenges 嗜热酸性绿藻的全细胞生物技术应用：机遇与挑战

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-19 DOI: 10.1016/j.procbio.2025.10.008

Longinus Ifeanyi Igbojionu , Marta Maso Martinez , Alan D. Goddard , Alfred Fernandez-Castane

Extremophiles are microorganisms that thrive in harsh environmental conditions where no life exists. These environmental conditions can involve temperature, acidity, salinity, pressure, or radiation that are typically uninhabitable to most microorganisms. The thermoacidophilic Archaea (e.g., Acidianus, Metallosphaera and Sulfolobus) can thrive at high temperatures and low pHs. These organisms are capable of autotrophic, lithotrophic, heterotrophic, chemoheterotrophic, and chemolithoautotrophic lifestyles, and thus can easily be cultivated on many different substrates. Additionally, their innate capacities to oxidise ferrous iron and/or reduced inorganic sulfur compounds have gained recognition for their utility in biomining operations. Members of Acidianus and Metallosphaera have been applied in bioleaching operations to extract valuable metals from low-grade ores and mineral concentrates. The advances in Sulfolobus genetics have presented opportunities for their application as platform organisms in biotechnological and biorefinery processes due to the ability to cultivate these extremophilic organisms under non-sterile conditions. Furthermore, utilising inexpensive and sustainable feedstock such as lignocellulosic biomass is one key advantage of lowering cultivation costs. This review presents current research developments on thermoacidophilic Sulfolobales members, emphasising their whole-cell applications for biomining operations, biotechnology and biorefinery. Several laboratory-scale studies found these organisms promising for large-scale deployment based on their unique characteristics.

极端微生物是在没有生命存在的恶劣环境条件下茁壮成长的微生物。这些环境条件可能包括温度、酸度、盐度、压力或辐射，这些条件通常不适合大多数微生物居住。嗜热酸性古生菌（如Acidianus， Metallosphaera和Sulfolobus）可以在高温和低ph下繁殖。这些生物能够自养、养石、异养、化养异养和化养养自养的生活方式，因此可以很容易地在许多不同的基质上培养。此外，它们固有的氧化亚铁和/或还原无机硫化合物的能力已因其在生物矿化操作中的用途而得到认可。酸化矿和金属矿的成员已应用于生物浸出作业，从低品位矿石和矿物精矿中提取有价金属。由于能够在非无菌条件下培养这些极端微生物，因此在亚硫酸盐遗传学方面的进展为其作为生物技术和生物炼制过程中的平台生物提供了机会。此外，利用廉价和可持续的原料，如木质纤维素生物质，是降低种植成本的一个关键优势。本文综述了嗜热酸性亚砜类化合物的最新研究进展，重点介绍了它们在生物矿化操作、生物技术和生物炼制中的全细胞应用。几项实验室规模的研究发现，基于这些生物的独特特征，它们有望大规模部署。

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引用次数: 0

Enhancing Wolffia globosa cultivation: Feasibility of aerated suspended-growth systems as alternative cultivating option and the influence of abiotic factors on biomass production, CO₂ capture, and biochemical composition 加强全球沃尔夫的培养：曝气悬浮生长系统作为替代培养方案的可行性以及非生物因素对生物量生产、CO 2捕获和生化组成的影响

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-15 DOI: 10.1016/j.procbio.2025.10.006

Kittikoon Sucunthowong , Shiva Rezaei Motlagh , Jareeluk Plengsakul , Sorawit Powtongsook , Kasidit Nootong

This study evaluated the feasibility of well-mixed suspended-growth systems for cultivating Wolffia globosa and investigated the effects of abiotic factors on growth, CO₂ capture, and biochemical composition of Wolffia. Continuous aeration at 0.4 and 0.8 vvm significantly improved biomass productivity compared to static water cultivation systems, while continuous stirring at 600 rpm caused extensive frond damage and lower biomass concentrations. Under optimized aerated conditions (500 mg/L initial biomass, 80 mg N/L initial nitrate, and 402 μmol photon/m²·s light intensity), the maximum biomass productivity reached 199.94 ± 25.88 mg/L·d, representing 83–277 % increase over static water cultivation. The corresponding CO₂ fixation rate (283.71 ± 36.73 mg CO₂/L·d) was comparable to those of Chlorella, highlighting the potential of W. globosa for biological CO₂ mitigation. Protein contents under optimized condition was 21.5 % of dry weight, with nitrate availability positively influencing protein accumulation. Key carotenoids, including lutein, β-carotene, neoxanthin, and violaxanthin, were detected, with no significant difference (p > 0.05) between suspended growth and static water systems. Furthermore, due to large frond size, biomass harvesting can be effectively conducted using filtration with low-cost, locally available materials.

本研究评估了混合悬浮体系培养全球狼尾草的可行性，并研究了非生物因素对狼尾草生长、CO2捕获和生化组成的影响。与静态水培养系统相比，0.4和0.8 vvm的连续曝气显著提高了生物量生产力，而600 rpm的连续搅拌造成了广泛的叶片损伤和较低的生物量浓度。在初始生物量500 mg/L、初始硝酸盐80 mg N/L、光照强度402 μmol光子/m2·s的优化条件下，最大生物量生产力达到199.94 ± 25.88 mg/L·d，比静态水栽培提高83 ~ 277 %。相应的CO2固定率（283.71 ± 36.73 mg CO2/L·d）与小球藻相当，凸显了小球藻在生物CO2减缓方面的潜力。优化条件下蛋白质含量为干重的21.5 %，硝态氮有效性对蛋白质积累有正向影响。关键类胡萝卜素，包括叶黄素、β-胡萝卜素、新黄质和紫黄质，在悬浮生长和静态水系统之间无显著差异（p >； 0.05）。此外，由于大叶尺寸，生物质收集可以有效地进行过滤与低成本，当地可用的材料。

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引用次数: 0

Extraction of proteins from streptomycin fermentation residue using aminosilane-modified magnetic Fe3O4 nanoparticles immobilized with neutral protease and lysozyme 中性蛋白酶和溶菌酶固定化氨基硅烷修饰磁性Fe3O4纳米颗粒提取链霉素发酵渣中的蛋白质

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-14 DOI: 10.1016/j.procbio.2025.10.001

Zaixing Li , Wenjing Zhang , Wei Gao , Rui Liu , Shangyi Shu , Bingbing Xu , Xue Qin , Chen Hong , Nannan Wang

Streptomycin fermentation residues (SFR) are rich in protein, and enzyme hydrolysis is an efficient technique for recovering protein from these residues. This study explores the extraction of protein from SFR using immobilized enzyme technology, aiming to achieve efficient protein extraction and repeated enzyme reuse, thereby promoting the cost-effective and efficient recovery of proteins from antibiotic fermentation residues. In this experiment, we synthesized amino-silane modified magnetic Fe₃O₄ nanoparticles (Fe₃O₄@SiO₂-NH₂) and used them as carriers to immobilize lysozyme (Lys) and neutral protease (Neu) on the surface of Fe₃O₄@SiO₂-NH₂ (CE-Fe₃O₄@SiO₂-NH₂), successfully preparing a recyclable biocatalyst. This catalyst was then used for protein extraction from SFR. In a 20.0 % glutaraldehyde solution, the composite enzyme (CE) concentration was 0.6 mg·mL⁻¹, and the crosslinking time was 5 h, resulting in the highest immobilization efficiency (74.7 % ± 0.48 %) and enzyme relative activity (89.2 % ± 0.59 %). The CE-Fe₃O₄@SiO₂-NH₂ still exhibited superparamagnetic properties, with a saturation magnetization of 54 emu·g⁻¹. After 30 days of storage, the CE-Fe₃O₄@SiO₂-NH₂ retained 75.6 % ± 1.12 % of its initial activity. Under extraction conditions of pH 6.0, 45℃, and 3 h, CE-Fe₃O₄@SiO₂-NH₂ successfully extracted 59.3 % ± 1.08 % of the protein from SFR. Furthermore, after five cycles of reuse, CE-Fe₃O₄@SiO₂-NH₂ maintained 30.8 % ± 1.20 % of its protein extraction capacity. The catalyst demonstrated significant protein extraction efficiency from SFR and can be repeatedly recovered, improving processing efficiency, reducing costs, and enhancing the practical application value of the enzyme.

链霉素发酵残留物含有丰富的蛋白质，酶法是一种从这些残留物中回收蛋白质的有效技术。本研究探索利用固定化酶技术从SFR中提取蛋白质，旨在实现蛋白质的高效提取和酶的重复再利用，从而促进抗生素发酵残留物中蛋白质的经济高效回收。在本实验中，我们合成了氨基硅烷修饰的磁性Fe3O4纳米颗粒（Fe3O4@SiO2-NH2），并将其作为载体固定在Fe3O4@SiO2-NH2 （CE-Fe3O4@SiO2-NH2）表面的溶菌酶（Lys）和中性蛋白酶（Neu），成功制备了可回收的生物催化剂。然后用该催化剂从SFR中提取蛋白质。在20.0 %戊二醛溶液中，复合酶（CE）浓度为0.6 mg·mL−1，交联时间为5 h，获得了最高的固定化效率（74.7 %±0.48 %）和酶相对活性（89.2 %±0.59 %）。CE-Fe3O4@SiO2-NH2仍然表现出超顺磁性，饱和磁化强度为54 emu·g−1。30天后，CE-Fe3O4@SiO2-NH2保留了其初始活性的75.6% %±1.12 %。在pH 6.0、45℃、3 h的提取条件下，CE-Fe3O4@SiO2-NH2成功地从SFR中提取了59.3 %±1.08 %的蛋白。此外，在重复使用5次后，CE-Fe3O4@SiO2-NH2的蛋白质提取率保持在30.8 %±1.20 %。该催化剂从SFR中提取蛋白质的效率显著，且可重复回收，提高了处理效率，降低了成本，增强了酶的实际应用价值。

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引用次数: 0

Evaluating propionate production in Clostridium ljungdahlii inoculated bioelectrochemical system 永达梭菌接种生物电化学体系丙酸产量的评价

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-13 DOI: 10.1016/j.procbio.2025.10.003

Zhen Fang , Qichao Fan , Yu Huang , Beisong Xu , Zhi-Jie Yang , Yang Gu , Vinod Kumar , Yang-Chun Yong

Bioproduction of chemical building block such as propionic acid (propionate) is of great interest in current time as it is a sustainable alternative to petrochemical synthesis. Clostridium ljungdahlii as an electroactive homoacetogen is able to ferment sugars or utilize CO₂ to produce organic acids under bioelectrochemical system (BES). However, few reports evaluated the ability of bioelectrochemically assisted propionate production via C. ljungdahlii. In this study, BES equipped with three-dimensional nanostructure electrode and inoculated with C. ljungdahlii in cathode chamber was developed to evaluate propionate production. It found that applied negative potential on cathode was necessary to stimulate propionate production from fructose. Bioelectrochemical characterizations of bacteria attached electrode revealed that C. ljungdahlii was able to assimilate extracellular electron. Notably, optimization of negative potential on electrode remarkably increased NAD(P)H/NAD(P)⁺ ratio, ATP amounts, and propionate production. Furthermore, BES optimization results showed that propionate achieved 16.29 ± 0.16 mM maximum titer (43 % ratio of total organic acids) and 0.2 g/g mass yield (0.24 mol/mol carbon atom yield) from fructose. A theoretical value (1.6 mmol) of fixed CO₂ was calculated from mass/electron balance analysis. This work not only evaluated the performance of bioelectrochemical technology but also provided a new choice for sustainable and efficient propionate production.

生物生产化学原料如丙酸（丙酸酯）是目前人们非常感兴趣的，因为它是石化合成的可持续替代品。在生物电化学系统（BES）下，ljungdahlii梭状芽孢杆菌（Clostridium ljungdahlii）作为一种具有电活性的均质乙基菌，能够发酵糖类或利用CO2生产有机酸。然而，很少有报道评价生物电化学辅助丙酸盐生产的能力。本研究采用三维纳米结构电极，并在阴极室中接种永达氏弧菌，利用BES对丙酸酯的产率进行评价。研究发现，在阴极上施加负电位是刺激果糖生成丙酸的必要条件。细菌附着电极的生物电化学表征表明，永达菌具有吸收胞外电子的能力。值得注意的是，电极上的负电位优化显著提高了NAD(P)H/NAD(P)+比值、ATP量和丙酸产量。此外，BES优化结果表明，丙酸酯从果糖中获得的最大滴度为16.29 ± 0.16 mM（总有机酸比为43 %）和0.2 g/g质量收率（0.24 mol/mol碳原子收率）。通过质量/电子平衡分析计算出固定CO2的理论值（1.6 mmol）。本研究不仅评价了生物电化学技术的性能，而且为可持续高效生产丙酸盐提供了新的选择。

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引用次数: 0

Preparation of High Fischer ratio oligopeptides by the strategy of COF-TpPa-1 coated magnetic graphene oxide immobilized α-chymotrypsin and phenylalanine modified chitosan resin COF-TpPa-1包被磁性氧化石墨烯固定α-凝乳胰蛋白酶和苯丙氨酸修饰壳聚糖树脂制备高菲舍尔比寡肽

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-10 DOI: 10.1016/j.procbio.2025.10.002

Xueli Shi , Liting Shi , Yi Zhang , Yongxi He , Lingjiao Wang , Xuecheng Liu , Yonghong Zhou , Haoshuang Ju , Huihui Liu , Xiaolan Liu , Xiaojiao Sun

High Fischer Ratio Oligopeptides (HFO) are bioactive compounds with health benefits and applications in nutrition and therapeutics. However, high cost in enzymatic hydrolysis and non-specific adsorption losses of branched chain amino acids (BCAAs) posed challenges to large-scale preparation of HFO. This study introduced an innovative approach involving immobilization of α-chymotrypsin on COF-coated magnetic graphene (MG@TpPa-1-α-CT) for the hydrolysis of whey protein, coupled with using phenylalanine-modified chitosan resin (PCCR) as specific adsorbent for aromatic amino acids (AAAs). MG@TpPa-1 demonstrated an impressive immobilized-capacity of 144 mg·g⁻¹ for α-CT, along with a Kₘ of 18.98 mg·mL⁻¹ and a Vₘₐₓ of 13.44 µmol·min⁻¹ and exhibited enhanced working stability. Leveraging n-π interactions, PCCR demonstrated exceptional adsorption capabilities of 30.2 mg·g⁻¹ and 8.2 mg·g⁻¹ for phenylalanine and leucine, and elevated the F-value of whey protein hydrolysate to 28.2. This study provided a novel strategy for HFO’s large-scale preparation.

高菲舍尔比寡肽（HFO）是一种具有健康益处的生物活性化合物，在营养和治疗方面有着广泛的应用。然而，支链氨基酸（BCAAs）的高酶解成本和非特异性吸附损失给HFO的大规模制备带来了挑战。本研究提出了一种创新的方法，将α-凝乳胰蛋白酶固定在cof包被的磁性石墨烯（MG@TpPa-1-α-CT）上，用于水解乳清蛋白，并使用苯基丙氨酸修饰的壳聚糖树脂（PCCR）作为芳香氨基酸（AAAs）的特异性吸附剂。MG@TpPa-1对α-CT的固定容量为144 mg·g−1，K值为18.98 mg·mL−1，V值为13.44µmol·min−1，具有良好的稳定性。利用n-π相互作用，PCCR对苯丙氨酸和亮氨酸的吸附能力分别为30.2 mg·g−1和8.2 mg·g−1，并将乳清蛋白水解产物的f值提高到28.2。本研究为HFO的大规模制备提供了一种新的策略。

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引用次数: 0

Influence of type of organic solid waste and process temperature on lactate-driven hydrogen production and microbial community by self-fermentation 有机固体废弃物类型和工艺温度对乳酸产氢及微生物群落的影响

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-10-10 DOI: 10.1016/j.procbio.2025.10.004

Marisol Pérez-Rangel, Edith Villanueva-Galindo, Idania Valdez-Vazquez, Iván Moreno-Andrade

Organic solid waste (OSW), depending on its origin, shows differences in the content of carbohydrates, lipids, and proteins, influencing its potential for valorization. This study aimed to evaluate the influence of three types of OSW (restaurant food waste (RFW), market waste (MW), and the organic fraction of municipal solid waste (OFMSW)) and temperature (37 and 55°C) on hydrogen production, metabolites production, and microbial community composition. The native microbiota of each OSW served as inoculum. Results confirmed that the type of feedstock significantly influenced lactate-based hydrogen production, which was temperature-dependent. Mesophilic self-fermentation of OFMSW yielded the highest hydrogen production, reaching 257 mL H₂/L, 23 % higher than the obtained at thermophilic conditions and 71 % higher than that obtained from MW at mesophilic conditions. Hydrogen production of OFMSW at both 37°C and 55°C and MW at 37°C was correlated with the butyric acid production and the presence of the Clostridium genus. In contrast, RFW at 37 and 55 °C was associated with lactic acid production and a higher abundance of Lactobacillus and Bacillus, respectively. Results demonstrated that OFMSW can be directly converted to hydrogen, while RFW produces lactic acid, which could serve as a precursor for lactate-based hydrogen production in a two-stage process.

有机固体废物（OSW）的来源不同，其碳水化合物、脂质和蛋白质的含量也不同，从而影响其增值潜力。本研究旨在评估三种类型的OSW（餐馆食物垃圾（RFW），市场垃圾（MW）和城市生活垃圾（OFMSW）的有机部分）和温度（37和55℃）对产氢，代谢物产生和微生物群落组成的影响。每个OSW的原生微生物群作为接种物。结果证实，原料类型显著影响乳酸基制氢，这是温度依赖的。OFMSW自发酵产氢量最高，达到257 mL H2/L，比亲热条件下的产氢量高23 %，比亲热条件下的产氢量高71 %。37°C和55°C条件下OFMSW的产氢量和37°C条件下的产氢量与丁酸产量和梭状芽孢杆菌属的存在相关。相比之下，37°C和55°C的RFW分别与乳酸产量和乳杆菌和芽孢杆菌的较高丰度有关。结果表明，OFMSW可以直接转化为氢，而RFW可以产生乳酸，乳酸可以作为两阶段制氢的前体。

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引用次数: 0

Bioconversion of phytosterols to 9-hydroxy-3-oxo-4-pregnene-20-carboxylic acid methyl ester by rerouting of phytosterol degradation pathways in Mycolicibacterium neoaurum 植物甾醇在新金黄色分枝杆菌中转化为9-羟基-3-氧-4-孕烯-20-羧酸甲酯

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-09-26 DOI: 10.1016/j.procbio.2025.09.012

Xinxin Chen , Xiaohan Jiang , Bo Zhang , Zhiqiang Liu , Yuguo Zheng

9-Hydroxy-3-oxo-4-pregnene-20-carboxylic acid methyl ester (9-OH-3-OPCM) is a representative C-22 steroid precursor in the synthesis of progestational based drug. However, 9-OH-3-OPCM production from phytosterol in Mycolicibacterium neoaurum (M. neoaurum) remains underdeveloped due to the complexity of the steroid metabolic pathway. In this study, we genetically modified the phytosterol metabolism in M. neoaurum and achieved an 85.9 % molar yield of 9-OH-3-OPCM with minimal by-products. Firstly, the biosynthesis of 9-OH-3-OPCM was achieved by disrupting ChsE1–2 and ChsH1–2 in KstDs-deficient M. neoaurum. Then, key enzymes encoded by ChsE4–5, Opccr, and SalA were inactivated to reduce the formation of by-products 9-hydroxy-3-oxo-4,17-pregadiene-20-carboxylic acid methyl ester (9-OH-3-OPDCM) and 9,21-dihydroxy-20-methyl-pregna-4-en-3-one (9-OH-4-HBC), as well as to increase 9-OH-3-OPCM production. Notably, multiple methyltransferases were overexpressed and screened to overcome suboptimal bioconversion from precursor 9-hydroxy-3-oxo-4-pregnene-20-carboxylic acid methyl ester (9-OH-3-OPC) to 9-OH-3-OPCM, which was attributed to inadequate methyltransferase activity. Overall, our findings demonstrate a strategy to improve the efficiency and purity of 9-OH-3-OPCM biosynthesis in M. neoaurum.

9-羟基-3-氧-4-孕烯-20-羧酸甲酯（9-OH-3-OPCM）是孕基药物合成中具有代表性的C-22甾体前体。然而，由于类固醇代谢途径的复杂性，新aurum分枝杆菌（M. neoaurum）从植物甾醇生产9-OH-3-OPCM仍然不发达。在这项研究中，我们对新aurum的植物甾醇代谢进行了基因改造，获得了85.9 %的9-OH-3-OPCM的摩尔产率，并且副产物最少。首先，通过破坏缺乏kstds的新金鼠的ChsE1-2和ChsH1-2，实现了9-OH-3-OPCM的生物合成。然后，失活ChsE4-5、Opccr和SalA编码的关键酶，减少副产物9-羟基-3-氧-4,17-预二烯-20-羧酸甲酯（9- oh -3- opdcm）和9,21-二羟基-20-甲基-孕-4-en-3-one （9- oh -4- hbc）的形成，增加9- oh -3- opcm的产量。值得注意的是，多个甲基转移酶被过度表达和筛选，以克服从前体9-羟基-3-氧-4-孕烯-20-羧酸甲酯（9-OH-3-OPC）到9-OH-3-OPCM的次优生物转化，这是由于甲基转移酶活性不足造成的。总的来说，我们的研究结果表明了一种提高新aurum中9-OH-3-OPCM生物合成效率和纯度的策略。

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引用次数: 0

Deep learning guided mutagenesis of signal peptide in Escherichia coli 深度学习引导大肠杆菌信号肽诱变

IF 4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry

Pub Date : 2025-09-25 DOI: 10.1016/j.procbio.2025.09.011

Ruei-En Hu, Hsueh-Jui Liu, Hui-Chun Chen, I-Son Ng

Signal peptides (SPs) play critical roles in protein secretion, yet their application in bioengineering remains limited by context-dependent functionality, host specificity, and genetic stability. This study presents a rational mutagenesis strategy for signal peptides in Escherichia coli under deep learning model and identifies the mutation-tolerant variants. At first, we generated 438 signal peptide sequences using SignalP 6.0. Through systematic modification of N-terminal, hydrophobic regions (H-region), and C-terminal sequence of signal peptide, we retrieved the high mutation tolerance existing in H-region. Among seven high mutation-tolerant signal peptides, SP22_54 from YjdP protein demonstrated exceptional robustness, maintaining functionality after extensive mutations. Further analysis revealed that functional SP mutants consistently preserved hydrophobic amino acid content with a minimal charge property. The SP_r1 mutant effectively mediated secretion of human carbonic anhydrase II (hCAII). The identified sequence determinants of mutation tolerance provide valuable insights for rational signal peptide design in biotechnological applications, enhancing reliability and cost-effectiveness of recombinant protein production across diverse expression systems.

信号肽（SPs）在蛋白质分泌中起着至关重要的作用，但其在生物工程中的应用仍然受到环境依赖功能、宿主特异性和遗传稳定性的限制。本研究提出了一种基于深度学习模型的大肠杆菌信号肽的合理诱变策略，并识别出耐突变变异体。首先，我们使用SignalP 6.0生成了438个信号肽序列。通过对信号肽的n端、疏水区（h区）和c端序列进行系统修饰，我们恢复了h区存在的高突变耐受性。在7个高耐突变信号肽中，来自YjdP蛋白的SP22_54表现出优异的稳健性，在广泛突变后仍能保持功能。进一步分析表明，功能性SP突变体始终保持疏水氨基酸含量，并具有最小电荷特性。SP_r1突变体有效介导人碳酸酐酶II （hCAII）的分泌。已确定的突变耐受性序列决定因素为生物技术应用中的合理信号肽设计提供了有价值的见解，提高了跨不同表达系统的重组蛋白生产的可靠性和成本效益。

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引用次数: 0