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The Biosynthesis of the Monoterpene Tricyclene in E. coli through the Appropriate Truncation of Plant Transit Peptides 通过适当截短植物转运肽在大肠杆菌中生物合成单萜三环烯
Pub Date : 2024-03-20 DOI: 10.3390/fermentation10030173
Meijia Zhao, Shaoheng Bao, Jiajia Liu, Fuli Wang, G. Yao, Penggang Han, Xiukun Wan, Chang Chen, Hui Jiang, Xinghua Zhang, Wenchao Zhu
Tricyclene, a tricyclic monoterpene naturally occurring in plant essential oils, holds potential for the development of medicinal and fuel applications. In this study, we successfully synthesized tricyclene in E. coli by introducing the heterologous mevalonate (MVA) pathway along with Abies grandis geranyl diphosphate synthase (GPPS) and Nicotiana sylvestris tricyclene synthase (TS) XP_009791411. Initially, the shake-flask fermentation at 30 ◦C yielded a tricyclene titer of 0.060 mg/L. By increasing the copy number of the TS-coding gene, we achieved a titer of 0.103 mg/L. To further enhance tricyclene production, optimal truncation in the N-terminal region of TS XP_009791411 resulted in an impressive highest titer of 47.671 mg/L, approximately a 794.5-fold improvement compared to its wild-type counterpart. To the best of our knowledge, this is the highest titer of the heterologous synthesis of tricyclene in E. coli. The SDS-PAGE analysis revealed that lowering induction temperature and truncating the random coil N-terminal region effectively improved TS solubility, which was closely associated with tricyclene production levels. Furthermore, by truncating other TSs, the titers of tricyclene were improved to different degrees.
三环烯是一种天然存在于植物精油中的三环单萜,具有开发药用和燃料应用的潜力。在本研究中,我们通过引入异源甲羟戊酸(MVA)途径以及大叶悬铃木香叶酯二磷酸合成酶(GPPS)和烟草三环烯合成酶(TS)XP_009791411,在大肠杆菌中成功合成了三环烯。最初,在 30 ◦C 下进行摇瓶发酵产生的三环烯滴度为 0.060 mg/L。通过增加 TS 编码基因的拷贝数,我们获得了 0.103 mg/L 的滴度。为了进一步提高三环烯的产量,我们对 TS XP_009791411 的 N 端区域进行了最佳截断,其最高滴度达到了令人印象深刻的 47.671 mg/L,与野生型相比提高了约 794.5 倍。据我们所知,这是大肠杆菌异源合成三环烯的最高滴度。SDS-PAGE 分析表明,降低诱导温度和截短随机线圈 N 端区域可有效提高 TS 的溶解度,这与三环烯的生产水平密切相关。此外,通过截短其他 TS,三环烯的滴度也有不同程度的提高。
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引用次数: 0
Yeast Biotechnology 6.0 酵母生物技术 6.0
Pub Date : 2024-03-19 DOI: 10.3390/fermentation10030172
R. Willaert
This Special Issue continues the “Yeast Biotechnology” Special Issue series of the MDPI journal Fermentation [...]
本特刊是 MDPI 期刊《发酵》"酵母生物技术 "特刊系列的延续 [...]
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引用次数: 0
Correction: Bintsis, T.; Papademas, P. The Evolution of Fermented Milks, from Artisanal to Industrial Products: A Critical Review. Fermentation 2022, 8, 679 更正:Bintsis, T.; Papademas, P. The Evolution of Fermented Milks, from Artisanal to Industrial Products:A Critical Review.Fermentation 2022, 8, 679
Pub Date : 2024-03-19 DOI: 10.3390/fermentation10030171
T. Bintsis, P. Papademas
In the original publication [...]
在最初的出版物中 [...]
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引用次数: 0
Effect of High Altitude on Serum Biochemical Parameters, Immunoglobulins, and Rumen Metabolism of Sanhe Heifers 高海拔对三河小母牛血清生化指标、免疫球蛋白和瘤胃代谢的影响
Pub Date : 2024-03-18 DOI: 10.3390/fermentation10030170
Xinyu Zhang, Z. Cao, H. Yang, Yajing Wang, Wei Wang, Shengli Li
Rumen metabolism is closely related to feed utilization and the environmental adaptability of cows. However, information on the influence of altitude on ruminal metabolism is limited. Our study aimed to investigate differences in rumen metabolism and blood biochemical indicators among Sanhe heifers residing at various altitudes. A total of 20 serum and ruminal fluid samples were collected from Sanhe heifers in China, including those from Hulunbeier City (approximately 700 m altitude; 119°57′ E, 47°17′ N; named LA) and Lhasa City (approximately 3650 m altitude; 91°06′ E, 29°36′ N; named HA). Compared with LA heifers, HA heifers had higher levels of serum cortisol, glucose, and blood urea nitrogen (p < 0.05) and lower Ca2+ concentrations (p < 0.05). Using liquid chromatography–mass spectrometry (LC–MS)-based untargeted metabolomic technology, we identified a significant difference in 312 metabolites between the LA and HA groups. Metabolic pathway analysis, based on significantly different rumen metabolites, identified 20 enriched metabolic pathways within hierarchy III, which are encompassed within 6 broader metabolic pathways in hierarchy I. This study constitutes the first elucidation of the altitudinal adaptation mechanism of ruminants from the perspective of rumen metabolism, thereby offering a novel angle for investigating high-altitude adaptation in both humans and animals.
瘤胃代谢与饲料利用率和奶牛的环境适应能力密切相关。然而,有关海拔高度对瘤胃代谢影响的信息十分有限。我们的研究旨在调查居住在不同海拔地区的三河母牛瘤胃代谢和血液生化指标的差异。我们共采集了 20 头三河小母牛的血清和瘤胃液样本,其中包括来自呼伦贝尔市(海拔约 700 米,东经 119°57′,北纬 47°17′,命名为 LA)和拉萨市(海拔约 3650 米,东经 91°06′,北纬 29°36′,命名为 HA)的小母牛。与LA母牛相比,HA母牛的血清皮质醇、葡萄糖和血尿素氮水平更高(P < 0.05),Ca2+浓度更低(P < 0.05)。利用基于液相色谱-质谱(LC-MS)的非靶向代谢组学技术,我们发现 LA 组和 HA 组之间的 312 种代谢物存在显著差异。这项研究首次从瘤胃代谢的角度阐明了反刍动物的高海拔适应机制,从而为研究人类和动物的高海拔适应性提供了一个新的角度。
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引用次数: 0
Distinct Short-Term Response of Intracellular Amino Acids in Saccharomyces cerevisiae and Pichia pastoris to Oxidative and Reductive Stress 酿酒酵母和酿醋酵母细胞内氨基酸对氧化和还原压力的不同短期反应
Pub Date : 2024-03-15 DOI: 10.3390/fermentation10030166
Burcu Şirin Kaya, E. Nikerel
Despite being frequently encountered, the effect of oxidative or reductive stress on the intracellular metabolism and the response of the intracellular metabolome of yeasts is severely understudied. Non-conventional yeasts are attracting increasing attention due to their large substrate portfolio of non-canonical pathways as well as their production and secretion of proteins. To understand the effects of both stresses on yeast, the conventional model yeast S. cerevisiae and the non-conventional model yeast P. pastoris were perturbed with 5 mM of hydrogen peroxide for oxidative stress and 20 mM of dithiothreitol for reductive stress in well-defined chemostat cultures at a steady state, and fermentation profiles, intracellular amino acid levels, and intracellular glutathione levels were measured. Although stable profiles of extracellular metabolites were observed, significant changes were measured in intracellular amino acid levels within the first five minutes. Collectively, the amino acids ranged from 0.5 to 400 µmol/gDW, with the most significant increase upon the induction of oxidative stress being seen in cysteine (up to 90%) for S. cerevisiae and in aspartate (up to 80%) for P. pastoris. Upon the induction of reductive stress, asparagine nearly halves in S. cerevisiae, while tryptophan decreases by 60% in P. pastoris. By inspecting the time traces of each amino acid, possible mechanisms of pathway kinetics are speculated. This work furthers our understanding of the response of metabolism to oxidative stress in two model yeasts.
氧化或还原压力对酵母菌细胞内代谢的影响以及细胞内代谢组的反应尽管经常遇到,但研究却严重不足。非常规酵母因其非典型途径的大量底物组合以及蛋白质的生产和分泌而日益受到关注。为了了解这两种胁迫对酵母的影响,在稳定状态下,用 5 mM 过氧化氢(氧化胁迫)和 20 mM 二硫苏糖醇(还原胁迫)对常规模式酵母 S. cerevisiae 和非常规模式酵母 P. pastoris 进行了扰动,并测量了发酵曲线、细胞内氨基酸水平和细胞内谷胱甘肽水平。虽然观察到了稳定的细胞外代谢物曲线,但在最初的五分钟内,细胞内氨基酸水平发生了显著变化。总的来说,氨基酸含量范围在 0.5 至 400 µmol/gDW 之间,在诱导氧化应激时,半胱氨酸的增加最为显著(S. cerevisiae 达 90%),天门冬氨酸的增加最为显著(P. pastoris 达 80%)。在诱导还原应激时,S. cerevisiae 的天冬酰胺几乎减少一半,而 P. pastoris 的色氨酸则减少 60%。通过观察每种氨基酸的时间轨迹,我们推测了通路动力学的可能机制。这项工作进一步加深了我们对两种模式酵母的新陈代谢对氧化应激反应的理解。
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引用次数: 0
Research Progress in Understanding the Molecular Biology of Cordyceps militaris 了解冬虫夏草分子生物学的研究进展
Pub Date : 2024-03-15 DOI: 10.3390/fermentation10030167
Lihong Wang, Ganghua Li, Xueqin Tian, Yitong Shang, Huanhuan Yan, Lihua Yao, Zhihong Hu
Cordyceps militaris (C. militaris) is a valued medicinal fungus that can be traced back thousands of years in traditional Chinese medicine (TCM). Both TCM and modern scientific research have confirmed the positive effects of C. militaris on human health. In recent years, C. militaris has gained wide popularity; unfortunately, strains often degrade during cultivation, resulting in a decline in fruiting bodies and active components that negatively impacts the development of C. militaris in the health food and medicine industries. This review summarizes the current progresses in research on the genomic, transcriptomic, proteomic, and genetic manipulation of C. militaris and discusses its primary metabolites and strain degradation mechanisms. The current challenges and future prospects of C. militaris research are also discussed.
冬虫夏草(Cordyceps militaris,简称 C.militaris)是一种珍贵的药用真菌,在中国传统医学(中医)中可追溯到数千年前。中医和现代科学研究都证实了冬虫夏草对人体健康的积极作用。遗憾的是,菌种在栽培过程中经常会发生退化,导致子实体和有效成分减少,这对小米赤芝在保健食品和医药行业的发展造成了负面影响。这篇综述总结了目前关于小米草基因组、转录组、蛋白质组和遗传操作的研究进展,并讨论了小米草的主要代谢产物和菌株降解机制。此外,还讨论了目前军事草研究面临的挑战和未来前景。
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引用次数: 0
Sanitizers Used for Fungal Spoilage Control in Dry-Fermented Cured Meat Production 用于控制干发酵腌肉生产中真菌腐败的消毒剂
Pub Date : 2024-03-15 DOI: 10.3390/fermentation10030169
Sarah Silva, A. O. Bernardi, M. V. Garcia, Thais Nunes Bisello, Larissa Borstmann, M. Copetti
Contamination caused by fungi stands out as a significant microbiological issue in the food industry, particularly leading to premature spoilage across various food segments, including the dry-fermented meat industry. The emergence of undesired fungi on product surfaces results in substantial economic losses. Once microorganisms infiltrate the food, contamination ensues, and their subsequent proliferation can adversely impact the product’s appearance, odor, flavor, and texture. This, in turn, leads to consumer rejection and negatively affects the commercial brand. Additionally, concerns persist regarding the potential presence of mycotoxins in these products. Given the detrimental effects of spoilage fungi in the food industry, practices such as thorough cleaning and sanitization become crucial to prevent contamination and subsequent premature deterioration. These measures play a pivotal role in ensuring the quality and safety of food, while also extending the shelf life of products. This review delves into the advantages, disadvantages, and factors that may influence the efficacy of commonly used sanitizers in the dry-fermented cured meat industry, including substances like sodium hypochlorite, peracetic acid, and benzalkonium chloride.
真菌造成的污染是食品工业中一个重要的微生物问题,尤其会导致包括干发酵肉类在内的各种食品过早变质。产品表面出现不受欢迎的真菌会造成巨大的经济损失。一旦微生物渗入食品,污染就会随之而来,其随后的扩散会对产品的外观、气味、风味和质地造成不利影响。这反过来又会导致消费者拒绝食用,并对商业品牌造成负面影响。此外,人们对这些产品中可能存在的霉菌毒素也一直存在担忧。鉴于腐败真菌在食品工业中的有害影响,彻底清洁和消毒等做法对于防止污染和随后的过早变质至关重要。这些措施在确保食品质量和安全以及延长产品保质期方面发挥着关键作用。本综述深入探讨了干发酵腌肉行业常用消毒剂(包括次氯酸钠、过乙酸和苯扎氯铵等物质)的优缺点以及可能影响其功效的因素。
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引用次数: 0
Source-Separated Industrial Wastewater Is a Candidate for Biogas Production through Anaerobic Digestion 源头分离的工业废水可通过厌氧消化生产沼气
Pub Date : 2024-03-14 DOI: 10.3390/fermentation10030165
J. Elliott, Christian Krohn, Andrew S. Ball
Anaerobic digestion is a potential treatment for industrial wastewater that provides valuable end-products, including renewable energy (biogas). However, waste streams may be too variable, too dilute at high volumes, or missing key components for stable digestion; all factors that increase costs and operational difficulty, making optimisation crucial. Anaerobic digestion may benefit from process intensification, particularly the novel combination of high-strength source-separated wastewater to minimise volume, together with the use of biosolids biochar as a chemical and microbial stabiliser. This study investigates the stability, yield, and microbial community dynamics of the anaerobic digestion of source-separated industrial wastewater from a food manufacturer and a logistics company, using biosolids biochar as an additive, focusing on gas and volatile fatty acid (VFA) production, process stability, and the microbial community using bench-scale semi-continuous reactors at 30- and 45-day hydraulic retention time (HRT). While gas yields were lower than expected, stability was possible at high HRT. Methane production reached 0.24 and 0.43 L day−1 per litre reactor working volume at 30- and 45-day HRT, respectively, despite high VFA concentration, and was linked to the relative abundance of Methanosarcina in the microbial community. Interactions between substrate, VFA concentration, and the microbial community were observed. Biochar-assisted anaerobic digestion holds promise for the treatment of source-separated wastewater.
厌氧消化是一种潜在的工业废水处理方法,可提供宝贵的最终产品,包括可再生能源(沼气)。然而,废物流可能变化太大,在高容量时太稀释,或缺少稳定消化的关键成分;所有这些因素都会增加成本和操作难度,因此优化至关重要。厌氧消化可从工艺强化中获益,特别是将高强度源分离废水与生物固体生物炭作为化学和微生物稳定剂相结合,最大限度地减少体积。本研究调查了厌氧消化食品制造商和物流公司源头分离的工业废水的稳定性、产量和微生物群落动态,使用生物固体生物炭作为添加剂,重点关注气体和挥发性脂肪酸(VFA)的产量、工艺稳定性,以及在 30 天和 45 天水力停留时间(HRT)下使用台式半连续反应器的微生物群落。虽然气体产量低于预期,但在高 HRT 条件下仍能保持稳定。尽管 VFA 浓度较高,但在 30 天和 45 天的 HRT 条件下,每升反应器工作容积的甲烷产量分别达到 0.24 升/天和 0.43 升/天,这与微生物群落中 Methanosarcina 的相对丰度有关。基质、VFA 浓度和微生物群落之间存在相互作用。生物炭辅助厌氧消化技术有望处理源分离废水。
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引用次数: 0
The Application of the Ultrasound Technique in the Production of Rosé and Red Wines 超声波技术在桃红葡萄酒和红葡萄酒生产中的应用
Pub Date : 2024-03-14 DOI: 10.3390/fermentation10030164
V. Lizama, I. Alvarez, M. J. García-Esparza
The application of the ultrasound technique (US) in the production of rosé and red wines has demonstrated that the aromatic composition of rose wine can be affected and that it contributes to increasing the color of red wines without increasing the extraction of astringent tannins. The ultrasound treatment has favored the extraction of anthocyanins, which has had an impact on the increase in color density (C.D.) and has allowed greater color stability over time. Moreover, significant differences have been found between the two US systems applied, with continuous treatment being more effective in the extraction of phenolic compounds than pulsed treatment. The application system of the US also affects the aromatic composition of the wines. These results are of interest, as some esters have been described as important odorants in wines.
超声波技术(US)在桃红葡萄酒和红葡萄酒生产中的应用表明,玫瑰葡萄酒的芳香成分会受到影响,超声波技术有助于增加红葡萄酒的颜色,而不会增加涩味单宁的提取。超声波处理有利于花青素的萃取,这对颜色密度(C.D.)的增加有影响,并使颜色随着时间的推移更加稳定。此外,两种超声波处理系统之间也存在明显差异,连续处理比脉冲处理对酚类化合物的提取更有效。US 应用系统也会影响葡萄酒的芳香成分。这些结果很有意义,因为一些酯类物质被认为是葡萄酒中重要的气味物质。
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引用次数: 0
Life Cycle Assessment of Exopolysaccharides and Phycocyanin Production with Arthrospira platensis 利用节肢动物板蓝根生产外多糖和植物花青素的生命周期评估
Pub Date : 2024-03-13 DOI: 10.3390/fermentation10030163
Isadora Cogo Badan, Sun-Hwa Jung, Rickwinder Singh, V. Vivekanand, Justus Knappert, Cornelia Rauh, Christoph Lindenberger
In the pursuit of sustainable solutions for contemporary environmental challenges arising from the increasing global demand for energy, this study delves into the potential of cyanobacteria, specifically Arthrospira platensis (commonly known as “spirulina”), as a versatile resource. Employing a life cycle assessment (LCA) in accordance with the ISO 14044:2006 standard and employing both midpoint and endpoint indicators, the study comprehensively evaluates environmental impacts. The research explored a range of scenarios, specifically investigating variations in light intensity and harvesting volume. These investigations were carried out using a pilot-scale photobioreactor, specifically an airlift reactor system featuring a horizontal tubular downcomer. The primary focus is on extracting valuable compounds, namely exopolysaccharides and phycocyanin. It emphasized the extraction of value-added products and strategic integration with a biogas plant for process heat, contributing to developing a sustainable supply network and offering insights into environmentally conscious algae cultivation practices with implications for renewable energy and the production of valuable products. The results emphasize the project’s potential economic feasibility with minimal energy impact from by-product extraction. The environmental assessment identifies marine ecotoxicity and fossil resource depletion as principal impacts, predominantly influenced by upstreaming and harvesting stages. After conducting comparisons across various scenarios, it was found that cultivations under higher light intensities have a lower environmental impact than cultivations with low light supply. However, regardless of light intensity, processes with shorter harvesting cycles tend to have a smaller environmental impact compared to processes with longer harvesting cycles. Overall, this research contributes a nuanced and realistic perspective, fostering informed decision-making in sustainable algae cultivation practices, with implications for renewable energy and valuable compound production.
为寻求可持续的解决方案,应对全球能源需求日益增长所带来的当代环境挑战,本研究深入探讨了蓝藻,特别是节旋藻(俗称 "螺旋藻")作为一种多功能资源的潜力。该研究采用符合 ISO 14044:2006 标准的生命周期评估 (LCA),并采用中点和终点指标,全面评估了对环境的影响。研究探索了一系列方案,特别是调查了光照强度和收割量的变化。这些研究是利用中试规模的光生物反应器进行的,特别是采用水平管式导流器的空运反应器系统。主要重点是提取有价值的化合物,即外多糖和植物花青素。该项目强调提取高附加值产品,并与沼气厂进行战略整合,以获取工艺热量,这有助于开发可持续供应网络,并为具有环保意识的藻类培育方法提供了见解,对可再生能源和有价值产品的生产具有重要意义。评估结果强调了该项目的潜在经济可行性,并将副产品提取对能源的影响降至最低。环境评估认为,海洋生态毒性和化石资源枯竭是主要影响因素,主要受上游和收获阶段的影响。在对各种方案进行比较后发现,光照强度较高的栽培对环境的影响低于光照强度较低的栽培。然而,无论光照强度如何,收获周期较短的过程对环境的影响往往小于收获周期较长的过程。总之,这项研究提供了一个细致而现实的视角,有助于在可持续藻类培育实践中做出明智的决策,并对可再生能源和有价值化合物的生产产生影响。
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引用次数: 0
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Fermentation
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