Journal of bioscience and bioengineering最新文献_第7页

Reproducing in vitro artificial gut microbiota using glycerol stocks of fecal cultures combined with different prebiotic additives. 利用粪便培养物的甘油原液结合不同益生元添加剂进行体外人工肠道菌群的繁殖。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-09-01 Epub Date: 2025-07-01 DOI: 10.1016/j.jbiosc.2025.06.002

Kengo Sasaki, Yasunobu Takeshima, Ayami Fujino

Artificial human microbiota can be produced in gut simulators from cryopreserved stocks. They are used for in vitro fermentation models and as alternative material for fecal microbiota transplantation therapy. However, current methods have limited information on microbial structure at the genus level and present challenges during cryopreservation. In this study, we used an edible glycerol stock of fecal batch culture instead of fresh feces to create artificial gut microbiota. Three glycerol stocks, generated through in vitro fecal fermentation with different prebiotic additives (such as fructooligosaccharide, xylan, pectin, and guar gum), were combined. Profiling via 16S rRNA gene amplicon sequencing revealed that the artificial gut microbiota derived from the combined glycerol stocks showed more amplicon sequence variants than those from a single glycerol stock. In the artificial microbiota, relative abundance values of common genera such as Bifidobacterium, Bacteroides, Prevotella, Faecalibacterium, and Escherichia were more than 10 % of those found in the original feces. Other commensal genera such as Collinsella, Anaerobutyricum hallii (formerly Eubacterium hallii) group, Anaerostipes, Blautia, Dorea, Lachnospiraceae UCG-004, and Oscillospiraceae UCG-003 were similarly maintained. Our data indicated that combining glycerol stocks of fecal cultures with different additives in a batch-type gut simulator is a useful option for producing artificial gut microbiota, the taxonomic compositions of which are comparable to those of the original feces.

人工人体微生物群可以在肠道模拟器中从冷冻储存的库存中产生。它们用于体外发酵模型和作为粪便微生物群移植治疗的替代材料。然而，目前的方法在属水平上对微生物结构的信息有限，并且在冷冻保存过程中存在挑战。在本研究中，我们使用粪便分批培养的食用甘油原液代替新鲜粪便来制造人工肠道菌群。采用不同的益生元添加剂（如低聚果糖、木聚糖、果胶和瓜尔胶）体外粪便发酵生成的三种甘油原液进行组合。通过16S rRNA基因扩增子测序分析显示，来自组合甘油培养基的人工肠道微生物群比来自单一甘油培养基的人工肠道微生物群显示出更多的扩增子序列变异。在人工微生物群中，双歧杆菌、拟杆菌、普雷沃氏菌、粪杆菌、埃希氏菌等常见菌属的相对丰度值均超过了原始粪便中的10%。其他共生属如Collinsella、Anaerobutyricum hallii（原真杆菌hallii）组、Anaerostipes、Blautia、Dorea、Lachnospiraceae UCG-004和Oscillospiraceae UCG-003也得到了类似的维持。我们的数据表明，在间歇式肠道模拟器中，将粪便培养物的甘油存量与不同的添加剂组合是产生人工肠道微生物群的有用选择，其分类组成与原始粪便相当。

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引用次数: 0

High productivity of cellulase and xylanase enzymes in the mycelial-dispersed Pleurotus ostreatus Δpkac2 strain 菌丝分散平菇Δpkac2菌株纤维素酶和木聚糖酶高产性研究。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-29 DOI: 10.1016/j.jbiosc.2025.08.001

Yuitsu Otsuka , Moriyuki Kawauchi , Vladimir Elisashvili , Saori Endo , Kenya Tsuji , Akira Yoshimi , Chihiro Tanaka , Takehito Nakazawa , Toshikazu Irie , Yoichi Honda

White-rot fungi secrete unique enzymes to degrade plant cell wall components. These enzymes have the potential to improve the effective utilization of lignocellulosic biomass in a bio-based society. In our previous study, pkac2-disrupted strains of Pleurotus ostreatus were applied for high-density liquid culture by improving mycelial dispersibility. In this study, we investigated the productivity and transcriptional profiles of wood-degrading enzymes of the Δpkac2 strain in the high-density liquid cultivation. Cellulase and xylanase activities in the culture filtrate of Δpkac2 strains in liquid shaking culture were 7.8- and 49-fold higher than those of the wild-type (WT) strain, respectively. In this condition, the mycelial dry weight of Δpkac2 strains was two-fold higher than that of the WT, showing their greater efficiencies for cellulase and xylanase production than the WT. RNA-Seq analysis indicated that 9 of 35 predicted cellulase genes and two of five predicted xylanase genes were significantly upregulated in the Δpkac2 strains. On the contrary, the transcript levels of laccase-encoding genes were significantly decreased, indicating that pkac2 is involved in their transcriptional regulation. These results indicate that the Δpkac2 strains have high potential to produce cellulase and xylanase in the high-density liquid cultivation by increasing mycelial growth as well as upregulating the expression of relevant genes.

白腐真菌分泌独特的酶来降解植物细胞壁成分。在以生物为基础的社会中，这些酶具有提高木质纤维素生物质有效利用的潜力。在我们之前的研究中，通过提高菌丝的分散性，将pkac2破坏的平菇菌株用于高密度液体培养。在本研究中，我们研究了Δpkac2菌株在高密度液体培养下的木材降解酶的产量和转录谱。Δpkac2菌液摇培养滤液中纤维素酶和木聚糖酶活性分别比野生型（WT）高7.8倍和49倍。在此条件下，Δpkac2菌株的菌丝干重比WT高2倍，表明其生产纤维素酶和木聚糖酶的效率高于WT。RNA-Seq分析表明，Δpkac2菌株35个纤维素酶预测基因中有9个显著上调，5个木聚糖酶预测基因中有2个显著上调。相反，漆酶编码基因的转录水平显著降低，表明pkac2参与了它们的转录调控。上述结果表明，Δpkac2菌株在高密度液体培养条件下，通过促进菌丝生长和上调相关基因的表达，具有较高的生产纤维素酶和木聚糖酶的潜力。

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引用次数: 0

Injectability of temperature-responsive hydrogel derived from elastin-like polypeptide for cell delivery 由弹性蛋白样多肽衍生的温度响应水凝胶的可注射性。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-23 DOI: 10.1016/j.jbiosc.2025.08.003

Mutawakil Al Muqadasi , Keitaro Ii , Kei Nishida , Masayasu Mie , Eiry Kobatake

Injectable hydrogels are promising biomaterials for tissue engineering applications due to their ability to deliver bioactive compounds or cells with minimal invasiveness. Temperature-responsive in situ gelling hydrogels, which undergo transition from liquid to gel in response to temperature stimuli, are desirable candidates for injectable hydrogels. Elastin-like polypeptides (ELPs) are well-known temperature-responsive biomaterials for cell scaffolds, drug delivery, and tissue engineering, due to their biocompatibility, biodegradability, and tunable mechanical properties. However, due to high hydrophobicity and heterogeneous aggregation, the development of injectable hydrogel-derived ELPs remains limited. In our previous study, we designed coiled-coil unit-bound ELPs (CUBEs) hydrogel systems, which integrate ELPs, a polyaspartic acid (polyD) chain, a functional peptide, and a coiled-coil peptide. In this study, we evaluated the injectability and cell delivery potential of a basic CUBE hydrogel system, called O-CUBE (AVGVP)₄₂-D₈₈-CL. The O-CUBE protein solution was mixed with human cervical cancer (HeLa) cells, serving as a cell model, and subsequently injected into culture medium pre-warmed to 37 °C to initiate in situ gelation. O-CUBE protein was successfully gelled at an approximately 90 % gelation rate after injection at 37 °C within pH ranges of 6–8. Encapsulated HeLa cells exhibited spheroid morphology, indicating that the hydrogel facilitated cell–cell interactions in three-dimensional culture. Further evaluation using a DNA assay revealed that HeLa cells can survive and proliferate within the hydrogel. These results demonstrate that the CUBE hydrogel system is a promising candidate to deliver cells with minimal invasiveness.

可注射水凝胶是一种很有前途的生物材料，用于组织工程应用，因为它们能够以最小的侵入性递送生物活性化合物或细胞。温度响应型原位胶凝水凝胶在温度刺激下从液体转变为凝胶，是可注射水凝胶的理想候选物。弹性蛋白样多肽（ELPs）由于其生物相容性、可生物降解性和可调节的机械性能，是众所周知的温度响应生物材料，可用于细胞支架、药物输送和组织工程。然而，由于高疏水性和非均相聚集，可注射水凝胶衍生的elp的发展仍然有限。在我们之前的研究中，我们设计了螺旋状单位结合的ELPs （CUBEs）水凝胶体系，该体系整合了ELPs、聚天冬氨酸（polyD）链、功能肽和螺旋状肽。在这项研究中，我们评估了基本CUBE水凝胶体系的可注射性和细胞递送潜力，称为O-CUBE (AVGVP)42-D88-CL。将O-CUBE蛋白溶液与人宫颈癌（HeLa）细胞混合，作为细胞模型，随后注射到预热至37℃的培养基中进行原位凝胶化。O-CUBE蛋白在37°C下注射后，在6-8的pH范围内以约90%的凝胶率成功凝胶化。被包裹的HeLa细胞呈现球形形态，表明水凝胶在三维培养中促进了细胞间的相互作用。进一步的DNA分析表明，HeLa细胞可以在水凝胶中存活和增殖。这些结果表明，CUBE水凝胶系统是一种很有前途的候选系统，可以以最小的侵入性递送细胞。

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引用次数: 0

Identification of toluene degradation genes in Acinetobacter sp. Tol 5 不动杆菌Tol 5中甲苯降解基因的鉴定。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-21 DOI: 10.1016/j.jbiosc.2025.07.010

Shogo Yoshimoto , Maiko Hattori , Shori Inoue , Sakura Mori , Yuki Ohara , Katsutoshi Hori

Microbial degradation of aromatic compounds provides sustainable solutions for environmental remediation and bioconversion. Acinetobacter sp. Tol 5 is notable for its strong adhesiveness and potential as a biocatalyst for toluene degradation; however, its toluene metabolic pathway has not been fully elucidated. In this study, genomic analysis identified a cluster of genes in Tol 5 highly similar to the well-known tod operon of Pseudomonas putida, encoding enzymes responsible for toluene metabolism. Phylogenetic analyses indicated that these tod genes, unusual among Acinetobacter species, were likely acquired through horizontal gene transfer. Transcriptomic analyses revealed that todF and todC1 are co-transcribed, while the adjacent fadL2 gene, encoding a putative outer membrane transporter corresponding to P. putida todX, is independently transcribed. Growth experiments using gene-knockout mutants revealed that TodC1, the large subunit of dioxygenase, is essential for growth on toluene, whereas FadL2 is not essential. Growth curves on each carbon source further showed that the todC1 knockout mutant could metabolize benzoate, but not toluene or benzene, confirming that the TOD pathway is the primary route for toluene and benzene degradation in Tol 5. The identification of the functional TOD pathway, which is unique within Acinetobacter, provides genetic and biochemical insights for the development of Tol 5 as an efficient immobilized biocatalyst for the bioremediation and bioconversion of aromatic compounds.

芳香族化合物的微生物降解为环境修复和生物转化提供了可持续的解决方案。不动杆菌sp. Tol 5因其强大的粘附性和作为甲苯降解生物催化剂的潜力而闻名；然而，其甲苯代谢途径尚未完全阐明。在本研究中，基因组分析发现Tol 5中的一组基因与众所周知的恶臭假单胞菌tod操纵子高度相似，编码负责甲苯代谢的酶。系统发育分析表明，这些在不动杆菌物种中不常见的tod基因可能是通过水平基因转移获得的。转录组学分析显示todF和todC1是共转录的，而相邻的fadL2基因是独立转录的，fadL2基因编码一种推定的外膜转运蛋白，对应于恶臭假单胞菌todX。基因敲除突变体的生长实验表明，双加氧酶的大亚基TodC1是在甲苯上生长所必需的，而FadL2不是必需的。各碳源上的生长曲线进一步表明，todC1敲除突变体可以代谢苯甲酸酯，但不能代谢甲苯和苯，证实TOD途径是Tol 5中降解甲苯和苯的主要途径。TOD途径在不动杆菌中具有独特的功能，为Tol 5作为芳香族化合物生物修复和生物转化的高效固定化生物催化剂的开发提供了遗传学和生物化学方面的见解。

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引用次数: 0

Application of a mitigation method for nitrous oxide emission in a full-scale Carrousel reactor: Carbon footprint assessment 一种减缓氧化亚氮排放方法在全尺寸Carrousel反应堆中的应用：碳足迹评估。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-21 DOI: 10.1016/j.jbiosc.2025.07.009

Shohei Otomo , Akihiko Terada , Satoru Shibata , Tomoyuki Hori , Eisuke Tamura , Masahiro Ito , Yu-You Li , Fumiaki Takakai , Kunihiro Okano , Naoyuki Miyata , Shuhei Masuda

Nitrous oxide (N₂O) emissions from biological nitrogen removal processes in sewage treatment plants greatly contribute to their overall carbon footprint. The present study aimed to mitigate N₂O emissions from an elliptical Carrousel bioreactor in a full-scale plant. The oxygen supply agitators equipped at the influent point and the opposite side of the reactor was operated alternately. The dissolved N₂O (DN₂O) concentrations were lowered when the agitator at the influent point was suspended while that on the opposite side was running. This scenario was associated with high levels of complementary DNA (RNA) from potential complete denitrifying bacteria, indicating increased N₂O reduction activity utilizing influent organic matter. However, during periods of reduced influent organic load, dissolved oxygen (DO) levels temporarily increased; thereafter, DN₂O increased, accompanied by a decrease in DO. This fluctuation was associated with the accumulation of nitrite and nitrate resulting from ammonia oxidation during the high-DO periods. Based on these findings, an N₂O mitigation strategy was implemented: reducing the oxygen supply and increasing the running time of the opposite-side agitator during the low-organic-loading periods. This approach effectively decreased the DN₂O levels, although a certain degree of instability remained during rainfall events. The median N₂O emission factor decreased from 0.86 % to 0.28 %, reducing the annual carbon footprint of the plant by 14 %. This study provides valuable insights into N₂O mitigation for full-scale plants and demonstrates the great impact of N₂O reduction on their carbon footprint.

污水处理厂生物脱氮过程中产生的氧化亚氮（N2O）排放对其总体碳足迹有很大贡献。本研究旨在减少一氧化二氮排放从一个椭圆形的Carrousel生物反应器在一个完整的工厂。进水点和反应器对面的供氧搅拌器交替运行。当进水点的搅拌器悬停，而另一侧的搅拌器运行时，溶解的N2O （DN2O）浓度降低。这种情况与来自潜在完全反硝化细菌的高水平互补DNA （RNA）有关，表明利用进水有机物的N2O还原活性增加。然而，在降低进水有机负荷期间，溶解氧（DO）水平暂时升高；此后，DN2O升高，DO降低。这种波动与高do期氨氧化引起的亚硝酸盐和硝酸盐积累有关。基于这些发现，实施了一种N2O缓解策略：在低有机负荷期间减少氧气供应并增加对面搅拌器的运行时间。这种方法有效地降低了DN2O水平，尽管在降雨事件期间仍存在一定程度的不稳定性。N2O排放因子中位数从0.86%降至0.28%，使工厂年碳足迹减少14%。本研究为全面植物减少N2O提供了有价值的见解，并证明了N2O减少对其碳足迹的巨大影响。

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引用次数: 0

Nitrogen removal and N2O emissions in anammox reactors: Influence of reactor design on process performance and microbial communities 厌氧氨氧化反应器中的氮去除和N2O排放：反应器设计对工艺性能和微生物群落的影响。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-14 DOI: 10.1016/j.jbiosc.2025.07.007

Jean De Dieu Shema , Satoshi Nakai , Takehiko Gotoh , Wataru Nishijima , Toshikazu Suenaga

The assessment and mitigation of N₂O emissions from anammox-related processes is challenging for environmentally friendly wastewater treatment. This study evaluated the nitrogen removal efficiency (NRE), N₂O emissions, and microbial diversity in three laboratory-scale anammox reactors: a sequencing batch reactor (SBR) with a recirculation line, a continuous stirred tank reactor (CSTR) without a recirculation line (CSTR1), and a CSTR with a recirculation line (CSTR2). Across two operational phases with anammox biomass (dry weight) of 1.63 g L⁻¹ (phase I) and 5.44 g L⁻¹ (phase II), the SBR had a higher NRE and lower N₂O emissions than the CSTRs. The NREs in phase II were 70.7 ± 14.1 % for the SBR, 68.9 ± 15.7 % for CSTR2, and 41.9 ± 15.8 % for CSTR1. N₂O emissions from the SBR were reduced by 56 % in phase II relative to phase I. Microbial diversity declined, and community composition shifted during reactor operation. In phase II, the Shannon entropy indices were 4.77 (SBR), 4.61 (CSTR2), and 5.04 (CSTR1); higher diversity in CSTR1 correlated with lower anammox abundance and thus lower performance. Candidatus Jettenia caeni became the predominant anammox species. Gene analysis revealed a positive correlation between the abundance of anammox-specific 16S rRNA genes (targeted by Amx809f/Amx1066r) and NRE, while nirS and nirK gene copy numbers were inversely related to reactor performance (NRE and N₂O emissions). The copy numbers of nosZ genes (clade I and clade II) varied in phase II across different reactors, which potentially contributed to the differences in N₂O emission reductions observed during this phase.

评估和减少厌氧氨氧化相关工艺产生的N2O排放对环境友好型废水处理具有挑战性。本研究对3个实验室规模的厌氧氨氧化反应器进行了氮去除效率（NRE）、N2O排放和微生物多样性的评估，这3个反应器分别是：带循环线的序批式反应器（SBR）、不带循环线的连续搅拌槽式反应器（CSTR）（CSTR1）和带循环线的CSTR （CSTR2）。在厌氧氨氧化生物量（干重）分别为1.63 g L-1（一期）和5.44 g L-1（二期）的两个运行阶段，SBR的NRE高于cstr， N2O排放量低于cstr。二期NREs分别为SBR 70.7±14.1%、CSTR2 68.9±15.7%、CSTR1 41.9±15.8%。与第一阶段相比，第二阶段SBR的N2O排放量减少了56%，微生物多样性下降，群落组成在反应器运行过程中发生了变化。二期Shannon熵指数分别为4.77 （SBR）、4.61 （CSTR2）和5.04 (CSTR1)；CSTR1多样性越高，厌氧氨氧化丰度越低，生产性能越差。候选热氏菌（Candidatus Jettenia caeni）成为厌氧氧化菌的优势种。基因分析显示，厌氧氨氧化特异性16S rRNA基因（Amx809f/Amx1066r靶向）的丰度与NRE呈正相关，而nirS和nirK基因拷贝数与反应器性能（NRE和N2O排放）呈负相关。nosZ基因（进化枝I和进化枝II）的拷贝数在不同反应器的第二阶段有所不同，这可能导致该阶段观察到的N2O减排差异。

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引用次数: 0

Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation 海洋产油硅藻fisstulifera solaris基因组编辑技术的建立，以改善油脂积累。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-13 DOI: 10.1016/j.jbiosc.2025.07.008

Satoshi Murata , Natsuno Kushiyama , Yusuke Yabu , Kahori Watanabe , Taiga Fujii , Rein Yasui , Daisuke Nojima , Yoshiaki Maeda , Tomoko Yoshino , Yusuke Matsuda , Tsuyoshi Tanaka

Biofuel production using microalgae has attracted considerable attention owing to high growth rate and lipid accumulation properties. However, further enhancement in lipid productivity is required to render this economically feasible. CRISPR/Cas9, which is one of the powerful genome editing tools, is an essential technique that may solve this problem. The marine diatom Fistulifera solaris JPCC DA0580 is a promising candidate of the biofuel production, since it accumulates significant amount of lipids. However, genome editing techniques have not yet been established for F. solaris, which prevent the construction of valuable strains. In this study, CRISPR/Cas9-mediated specific gene knockout technique was established in F. solaris, through targeting adenine phosphoribosyl transferase gene (apt) and triacylglycerol (TAG) lipase gene (tgl1). Mutations in the target sequence were detected in apt- and tgl1-edited mutants. Moreover, the mutants showed distinct phenotypes, such as suppression of TAG degradation and resistance to 2-fluoroadenine. These results indicate the successful demonstration of CRISPR/Cas9-mediated genome editing in the oleaginous marine diatom F. solaris. Furthermore, oil degradation was successfully suppressed by knocking-out tgl1. The CRISPR/Cas9-mediated genome editing established in this study provides key molecular tools for both the basic biology and the future biotechnological applications of F. solaris, such as biofuel production.

利用微藻生产生物燃料因其高生长速度和油脂积累特性而受到广泛关注。然而，需要进一步提高脂质生产力，使其在经济上可行。CRISPR/Cas9是一种强大的基因组编辑工具，是解决这一问题的关键技术。海洋硅藻Fistulifera solaris JPCC DA0580是一种很有前途的生物燃料生产候选者，因为它积累了大量的脂质。然而，目前还没有针对solaris的基因组编辑技术，这阻碍了有价值菌株的构建。本研究通过靶向腺嘌呤磷酸核糖转移酶基因（apt）和三酰基甘油（TAG）脂肪酶基因（tgl1），建立了CRISPR/ cas9介导的solaris特异性基因敲除技术。在apt-和tgl1编辑的突变体中检测到靶序列的突变。此外，突变体表现出不同的表型，如抑制TAG降解和对2-氟腺嘌呤的抗性。这些结果表明，CRISPR/ cas9介导的基因组编辑在产油的海洋硅藻F. solaris中得到了成功的验证。此外，通过敲除tgl1成功抑制了原油降解。本研究建立的CRISPR/ cas9介导的基因组编辑为solaris的基础生物学和未来生物技术应用（如生物燃料生产）提供了关键的分子工具。

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引用次数: 0

Metabolomics-based investigation of the differences between traditional and modern soy sauce 基于代谢组学的传统酱油与现代酱油差异研究。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-11 DOI: 10.1016/j.jbiosc.2025.07.004

Yoshika Maekawa , Miki Nakamura , Miho Imamura , Eiichiro Fukusaki

Soy sauces can be classified into three categories based on the production method used: honjozo, kongojozo (mixed-brewing, amino acid-decomposed), and kongo (mixed, amino acid-decomposed) soy sauces. Although differences in flavor between traditional (honjozo) and modern (amino acid-decomposed) soy sauces are clear, knowledge of the differences in compound profiles and the relationship between these and the sensory characteristics that affect soy sauce quality is limited. Therefore, this study aimed to investigate the differences between traditional and modern soy sauce compounds using metabolomic analysis, and to investigate the compounds that may be correlated with differences in flavor. Non-targeted gas chromatography/mass spectrometry (GC/MS) metabolomics analysis was performed on nine traditional and six modern soy sauces to annotate 239 soy sauce compounds. Principal component analysis suggested that the production methods used formed clusters that affected the types and amounts of soy sauce compounds, and that the production method had a greater effect on soy sauce composition than did the aging barrels or type of soybean used. Traditional soy sauce was characterized by alcohols and esters, whereas modern soy sauce was characterized by pyrazines. A sensory evaluation revealed that traditional soy sauce was characterized by bitterness and astringency, whereas modern soy sauce was characterized by sweetness and viscosity, suggesting that the method of soy sauce production influences flavor differences. This is the first study to comprehensively characterize the effects of production methods, aging barrels, and soybean types on soy sauce compounds and how these compounds contribute to differences in flavor.

酱油根据制作方法可以分为honjozo、kongojozo（混合酿造、氨基酸分解）和kongo（混合、氨基酸分解）三大类。虽然传统酱油（本酒酱油）和现代酱油（氨基酸分解酱油）在风味上的差异是显而易见的，但对复合结构的差异以及这些差异与影响酱油质量的感官特征之间的关系的了解是有限的。因此，本研究旨在通过代谢组学分析来研究传统酱油和现代酱油化合物之间的差异，并研究可能与风味差异相关的化合物。对9种传统酱油和6种现代酱油进行了非靶向气相色谱/质谱（GC/MS）代谢组学分析，对239种酱油化合物进行了注释。主成分分析表明，不同的生产方式对酱油成分的类型和数量有一定的影响，生产方式对酱油成分的影响大于陈化桶或大豆类型。传统酱油以醇类和酯类为特征，现代酱油以吡嗪类为特征。感官评价结果表明，传统酱油以苦味和涩味为主，而现代酱油以甜味和粘性为主，说明酱油的制作方法影响了风味的差异。这是第一个全面描述生产方法、陈酿桶和大豆类型对酱油化合物的影响以及这些化合物如何导致风味差异的研究。

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引用次数: 0

Ribonucleoprotein-based CRISPR/Cas9 genome co-editing in Aspergillus luchuensis mut. kawachii 基于核糖核蛋白的CRISPR/Cas9基因组共编辑在葡曲霉中的应用kawachii。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-06 DOI: 10.1016/j.jbiosc.2025.07.006

Takefumi Karashima , Ken Oda , Taiki Futagami , Hideki Hokazono , Hideharu Takashita

In this study, we established a ribonucleoprotein-based clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome co-editing method for the white koji fungus, Aspergillus luchuensis mut. kawachii. To introduce the single guide RNA-Cas9 ribonucleoprotein complex into protoplast cells of A. luchuensis mut. kawachii, we investigated the conditions for protoplast preparation using Yatalase -Plus-. Subsequently, we employed the ribonucleoprotein-based method to knockout the ATP sulfurylase-encoding sC gene, which imparts selenate resistance in the model strain NBRC 4308 and the industrial strain No. 8046. Furthermore, we explored genome co-editing by simultaneously targeting sC along with either the orotidine 5′-phosphate decarboxylase-encoding pyrG gene or the transcriptional activator of protease genes-encoding prtR gene in NBRC 4308. The transformants were selected in medium containing selenate, resulting in the successful generation of pyrG- and prtR-knockout strains. Similarly, transformants were selected on medium containing selenate, resulting in the successful generation of prtR-knockout strain in No. 8046. These results demonstrate that the ribonucleoprotein-based genome co-editing method is applicable not only to the model strain but also to industrial strains, making it a promising approach for manipulating A. luchuensis mut. kawachii.

在本研究中，我们建立了一种基于核糖核蛋白簇状规则间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9 （Cas9）的白曲真菌Aspergillus luchuensis mut基因组共编辑方法。kawachii。目的：将单导RNA-Cas9核糖核蛋白复合物导入芦杉原生质体细胞。研究了利用Yatalase - plus -制备原生质体的条件。随后，我们采用基于核糖核蛋白的方法敲除了模型菌株NBRC 4308和工业菌株8046中编码ATP硫酰酶的sC基因，该基因赋予了硒酸盐抗性。此外，我们通过同时靶向sC与编码NBRC 4308的orotidine 5'-磷酸脱羧酶pyrG基因或编码prtR基因的蛋白酶基因转录激活因子，探索了基因组共编辑。在含硒酸盐的培养基中选择转化子，成功产生pyg -和prr -敲除菌株。同样，在含有硒酸盐的培养基上选择转化子，成功产生了编号8046的prr敲除菌株。这些结果表明，基于核糖核蛋白的基因组共编辑方法不仅适用于模式菌株，也适用于工业菌株，是一种很有前途的操作方法。kawachii。

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引用次数: 0

Enhanced heparosan biosynthesis in Escherichia coli Nissle 1917 through carbon flux redirection 通过碳通量重定向增强大肠杆菌Nissle 1917中肝磷脂的生物合成。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-08-05 DOI: 10.1016/j.jbiosc.2025.07.005

Fangqi Shao , Ruiji Wu , Zheng-Jun Li

Heparosan, a critical precursor for heparin production, is naturally biosynthesized as capsular polysaccharides by the probiotic strain Escherichia coli Nissle 1917 (EcN). This study presents a systematic metabolic engineering strategy to enhance heparosan biosynthesis through coordinated pathway engineering and carbon flux redirection. By disrupting glucose catabolism via deletion of zwf and pfkAB, we decoupled cell growth from heparosan synthesis while maintaining precursor availability, elevating titers from 137.68 mg/L to 422.11–486.13 mg/L in mixed carbon source cultures. Subsequent overexpression of UDP-glucose dehydrogenase, a key enzyme in UDP-glucuronic acid biosynthesis, achieved 1.04 g/L heparosan in shake-flask cultivations. Scale-up in a 5-L bioreactor demonstrated industrial scalability, yielding 4.34 g/L heparosan. Our work establishes EcN as a microbial chassis for glycosaminoglycan production and provides a generalizable framework for engineering complex polysaccharide biosynthesis through rational metabolic partitioning.

肝素聚糖是肝素生产的关键前体，是由大肠杆菌Nissle 1917 （EcN）益生菌天然合成的荚膜多糖。本研究提出了一种系统的代谢工程策略，通过协调的途径工程和碳通量重定向来促进肝磷脂的生物合成。通过删除zwf和pfkAB来破坏葡萄糖分解代谢，我们在保持前体可用性的同时，将细胞生长与肝磷脂合成分离，将混合碳源培养的滴度从137.68 mg/L提高到422.11-486.13 mg/L。随后，在摇瓶培养中，udp -葡萄糖醛酸生物合成的关键酶udp -葡萄糖脱氢酶的过表达量达到1.04 g/L。在5升生物反应器中进行放大，证明了工业可扩展性，产率为4.34 g/L。我们的工作建立了EcN作为糖胺聚糖生产的微生物基础，并通过合理的代谢分配为复杂多糖的工程生物合成提供了一个可推广的框架。

引用次数: 0