Journal of bioscience and bioengineering最新文献_第3页

Adaptive laboratory evolution optimizes an engineered phosphite utilization pathway in Synechococcus elongatus PCC 7942 适应性实验室进化优化了长聚球菌PCC 7942的工程亚磷酸酯利用途径。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-12-13 DOI: 10.1016/j.jbiosc.2025.11.006

Hiroki Murakami , Naoki Momokawa , Kei Motomura , Akio Kuroda , Ryuichi Hirota

Synthetic biology approaches enable the creation of promising chassis for practical application in various fields, though engineering of microbial metabolism often imposes a metabolic burden, potentially driving adaptive evolution during long-term cultivation. A previously established phosphite (Pt)-dependent metabolic system has proven to be an effective strategy for the containment of genetically engineered microorganisms, although its implementation accompanied a slight growth retardation. Here, we investigated the effect of long-term serial passaging cultivation on the Pt-dependent strain of Synechococcus elongatus PCC 7942, RH714. Compared with the originally constructed RH714, the passaged population of RH714 exhibited improved growth and a higher rate of Pt consumption in culture medium. Sequence analysis revealed point mutations within the introduced htxBCDE transporter genes, which are required for selective incorporation of Pt as a phosphorus nutrition. Introduction of the mutated gene cluster into S. elongatus PCC 7942 reproduced the traits of the passaged RH714 population, suggesting that these genetic changes enhance Pt transport activity and account for the observed phenotypes. Disruption of endogenous phosphate (Pi) transporter genes in the strains expressing the mutated htxBCDE-ptxD cluster abolished growth in Pi-containing medium, suggesting that the mutations in the transporter genes did not alter substrate specificity toward Pi. These results indicated that long-term passage cultivation developed an optimized mutant capable of efficient proliferation under the Pt metabolizing conditions without compromising its biocontainment capability.

尽管微生物代谢工程通常会带来代谢负担，但合成生物学方法能够为各种领域的实际应用创造有前途的基础，在长期培养过程中可能会推动适应性进化。先前建立的亚磷酸酯（Pt）依赖代谢系统已被证明是遏制基因工程微生物的有效策略，尽管其实施伴随着轻微的生长迟缓。本实验研究了长聚球菌（PCC） 7942， RH714对pt依赖菌株的长期连续传代培养的影响。与原构建的RH714相比，RH714的传代群体表现出更好的生长和更高的培养基Pt消耗率。序列分析显示，引入的htxBCDE转运体基因发生了点突变，这是Pt作为磷营养物选择性结合所必需的。将突变基因簇导入S. elongatus PCC 7942，可以再现传代的RH714群体的性状，表明这些遗传变化增强了Pt转运活性，并解释了所观察到的表型。在表达突变的htxBCDE-ptxD簇的菌株中，内源性磷酸盐（Pi）转运蛋白基因的破坏使其在含Pi的培养基中生长中断，这表明转运蛋白基因的突变并未改变对Pi的底物特异性。这些结果表明，长期传代培养产生了一个优化的突变体，能够在Pt代谢条件下高效增殖，而不影响其生物抑制能力。

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引用次数: 0

Development of dectin-1-binding peptides targeting dendritic cells for antigen delivery via ribosome display 通过核糖体展示靶向树突状细胞抗原递送的dectin-1结合肽的开发。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-12-07 DOI: 10.1016/j.jbiosc.2025.11.005

Yoshirou Kawaguchi , Md Shahin Sarker , Mina Yokoyama , Misuzu Nakaya , Takanatsu Hosokawa , Noriho Kamiya , Masahiro Goto

Dendritic cells (DCs) are professional antigen-presenting cells that play a central role in initiating and shaping adaptive immune responses. Targeting antigens to DCs has emerged as a promising strategy to enhance vaccine efficacy and tailor desired immune responses. While antibodies are well established as targeting molecules for DCs, the use of peptides remains underexplored despite their favorable tissue penetration and their ability to offer design flexibility. Here, we report the identification of dectin-1-binding peptides using a ribosome display-based in vitro directed evolution system. Dectin-1 is a C-type lectin receptor expressed on murine CD11b⁺ and human CD1c⁺ DCs, which plays a key role in antigen uptake and in directing immune responses toward specific pathways, particularly the Th17 pathway. Using recombinant murine dectin-1 as bait, we performed four rounds of ribosome display selection and obtained peptides with high affinity. Selected peptides fused to enhanced green fluorescent protein showed binding to recombinant dectin-1 and native dectin-1-expressing cells, including bone marrow-derived DCs. These results demonstrated the feasibility of peptide-based molecular targeting toward dectin-1⁺ DCs, which would not only provide a versatile platform for the development of next-generation vaccines and immunotherapies, but also a valuable tool for dissecting the functional roles of dectin-1⁺ DCs in immune regulation.

树突状细胞（dc）是一种专业的抗原呈递细胞，在启动和形成适应性免疫反应中起着核心作用。将抗原靶向dc已成为提高疫苗效力和定制所需免疫反应的一种有希望的策略。虽然抗体作为树突状细胞的靶向分子已经很好地建立起来，但尽管肽具有良好的组织穿透性和设计灵活性，但其使用仍未得到充分的探索。在这里，我们报告了使用基于核糖体显示的体外定向进化系统鉴定检测素-1结合肽。Dectin-1是一种c型凝集素受体，在小鼠CD11b+和人CD1c+ dc上表达，在抗原摄取和引导免疫反应向特定途径，特别是Th17途径中起关键作用。我们以重组小鼠dectin-1为诱饵，进行了四轮核糖体展示选择，获得了高亲和力的多肽。与增强绿色荧光蛋白融合的选定肽显示与重组dectin-1和天然表达dectin-1的细胞结合，包括骨髓来源的dc。这些结果证明了基于肽的分子靶向dectin-1+ dc的可行性，这不仅为下一代疫苗和免疫疗法的开发提供了一个通用的平台，而且为解剖dectin-1+ dc在免疫调节中的功能作用提供了有价值的工具。

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引用次数: 0

Recovering genomes from uncultured fungi with single-cell genomics 用单细胞基因组学从未培养的真菌中恢复基因组。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-12-05 DOI: 10.1016/j.jbiosc.2025.11.004

Nevin McCone , Masahito Hosokawa

Single-cell genomics (SCG) complements culture-independent metagenomics for accessing fungal genomes, particularly from lineages that remain uncultured. We contrast metagenomics, which excels when profiling community composition and metabolic potential but often underrepresents low-abundance fungi, with SCG, which first isolates individual cells or nuclei to generate single-amplified genomes (SAGs) and can recover rare or microdiverse taxa. We then organize existing fungal SCG applications into three subgroups: spore-level sequencing from host-enriched or environmental material; single-nucleus genomics for multinucleate fungi; and single-spore sequencing of haploid progeny for diploid linkage and chromosome phasing. Across studies, pooling and co-assembly of cognate cells improves completeness; key hurdles persist in wall lysis, whole-genome amplification bias, and contamination control. Practical advances include shallow sequencing for QC triage, nuclei pooling with normalized co-assembly, and hybrid long- and short-read assembly. SCG adds unique value where strain resolution and genotypic context matter, including host-to-mobile-element linkage, recovery of large biosynthetic gene clusters, and karyotype validation against telomere-to-telomere references. Used alongside metagenomics, SCG enables a strain-resolved view of fungal biodiversity and function, with incremental improvements across the SCG pipeline promising routine access to genomes from early-diverging and other environmentally embedded fungi.

单细胞基因组学（SCG）补充了独立于培养的宏基因组学，用于获取真菌基因组，特别是来自未培养的谱系。宏基因组学在分析群落组成和代谢潜力方面表现出色，但通常不代表低丰度真菌，而SCG则首先分离单个细胞或细胞核产生单扩增基因组（sag），并可以恢复稀有或微多样性的分类群。然后，我们将现有的真菌SCG应用分为三个亚组：从宿主富集或环境材料中进行孢子水平测序；多核真菌的单核基因组学研究单倍体后代的单孢子测序，用于二倍体连锁和染色体分期。在整个研究中，同源细胞的汇集和共同组装提高了完整性；关键的障碍仍然存在于细胞壁裂解、全基因组扩增偏倚和污染控制。实际的进展包括浅测序的QC分流，核池与标准化的共组装，和混合长和短读组装。SCG在菌株分辨率和基因型背景重要的情况下增加了独特的价值，包括宿主-移动元件链接，大型生物合成基因簇的恢复，以及针对端粒-端粒参考的核型验证。与宏基因组学一起使用，SCG可以实现真菌生物多样性和功能的菌株解析视图，随着SCG管道的逐步改进，有望常规获取早期分化和其他环境嵌入真菌的基因组。

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引用次数: 0

Zein/Fucoidan microcarriers promote myogenic differentiation via topographical cues and hydrodynamic modulation 玉米蛋白/岩藻聚糖微载体通过地形线索和流体动力学调节促进肌源性分化。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-12-04 DOI: 10.1016/j.jbiosc.2025.11.003

Wanli Xiong, Chengxin Ge, Botao Zhang, Ziying Chen, Yuzhe Guo, Qiaohui Lu, Wen-song Tan, Yan Zhou

Microcarrier surface topography and fluid shear stress (FSS) critically regulate cellular behavior. An edible zein/fucoidan microcarrier with 200 μm grooves was designed for this study. Computational fluid dynamics (CFD) simulations and experiments were combined to analyze surface microfluidic characteristics during dynamic culture and their cellular effects. The research demonstrates controlled myogenic differentiation through groove topography and FSS modulation for scalable cultured meat production. The results demonstrated that the grooved microcarriers exhibited excellent cell attachment and proliferation capacity in spinner flask dynamic culture, achieving a maximum cell density of 1.16 × 10⁶ cells/mL, comparable to commercial Cultispher-S microcarriers. The groove structure promoted cell alignment through contact guidance, significantly enhancing the gene expression of myogenic differentiation markers (myogenic differentiation 1, MyoD1; α-actinin; myosin heavy chain, MHC) and cell fusion (myomaker, MYMK). CFD simulations revealed that the grooves created a low-shear microenvironment (minimum average FSS: 3.93 × 10⁻² Pa, maximum: 1.18 × 10⁻¹ Pa), which effectively avoided high FSS-induced damage while maintaining mechanical stimulation. This optimal mechanical microenvironment further activated the expression of key genes involved in early-stage (MyoD1; myogenin, MyoG; myocyte enhancer factor 2C, MEF2C) and late-stage (α-actinin; myosin heavy chain 2, Myh2) myogenic differentiation. Flat and spherical microcarriers showed lower myogenic differentiation efficiency. This study elucidates the synergistic mechanism between groove structures and the low FSS microenvironment within grooves, providing a novel scaffold design rationale that combines biomimetic topology with fluid dynamics compatibility for large-scale cultured meat production.

微载体表面形貌和流体剪切应力（FSS）对细胞行为起着关键的调节作用。设计了一种具有200 μm凹槽的玉米蛋白/岩藻聚糖微载体。采用计算流体力学（CFD）模拟与实验相结合的方法，分析了动态培养过程中表面微流体的特性及其细胞效应。研究表明，通过沟槽地形和FSS调节控制的肌源性分化可用于规模化养殖肉生产。结果表明，微载体在旋转瓶动态培养中表现出良好的细胞附着和增殖能力，最大细胞密度为1.16 × 106个细胞/mL，与市售cultisper - s微载体相当。凹槽结构通过接触引导促进细胞排列，显著增强了成肌分化标志物（myogenic differentiation 1, MyoD1; α- actitin; myosin heavy chain， MHC）的基因表达和细胞融合（myomaker， MYMK）。CFD模拟表明，沟槽创造了一个低剪切微环境（最小平均FSS: 3.93 × 10-2 Pa，最大值：1.18 × 10-1 Pa），有效避免了高FSS引起的损伤，同时保持了机械刺激。这种优化的机械微环境进一步激活了参与早期（MyoD1; myogenin, MyoG; myocyte enhancer factor 2C, MEF2C）和晚期（α- actitin； myosin重链2，Myh2）成肌分化的关键基因的表达。扁平微载体和球形微载体的成肌分化效率较低。本研究阐明了沟槽结构与沟槽内低FSS微环境之间的协同机制，为大规模养殖肉类生产提供了一种结合仿生拓扑和流体动力学兼容性的新型支架设计原理。

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引用次数: 0

Spatial, temporal, and biotic changes in the natural abundances of stable carbon and nitrogen isotopes in a biological treatment reactor 生物处理反应器中稳定碳和氮同位素自然丰度的时空和生物变化。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-25 DOI: 10.1016/j.jbiosc.2025.11.002

Takashi Onodera , Kengo Kubota , Akinori Iguchi , Akihiro Nagamachi , Tadashi Tagawa , Gen Kanaya , Ayato Kohzu , Kazuaki Syutsubo

The natural abundances of stable isotopes were used to determine prey–predator relationships and material flows in a biological treatment reactor for municipal wastewater. The reactor used in this study was a down-flow hanging sponge (DHS), which is an alternative trickling filter that uses sponge as the packing material. The sponge retained sludge containing a wide variety of organisms, including microfauna. Stable isotope analysis revealed spatial, temporal, and biotic variations in the carbon and nitrogen stable isotope ratios (δ¹³C and δ¹⁵N) of the retained sludge and microfauna (water mites and fly larvae). In addition, adult flies and spiders were present and analyzed. The δ¹³C and δ¹⁵N in sludge were temporally and spatially similar along the reactor. The isotopic signature was associated with treatment characteristics such as a low nitrification efficiency in the DHS reactor. The δ¹³C and δ¹⁵N of sympatric fly larvae differed from those of water mites, which indicated dietary differences between the taxa. Interestingly, the water mites had higher δ¹³C and δ¹⁵N than the retained sludge, which indicated that they were in a higher trophic position in the food web. In addition, the δ¹³C and δ¹⁵N values of spiders were approximately 1 ‰–3‰ higher than those of adult flies. This strongly suggested that a prey–predator relationship existed between adult flies and spiders.

稳定同位素的天然丰度用于确定城市污水生物处理反应器中的捕食关系和物质流动。本研究采用的反应器为下流式悬挂海绵反应器（DHS），是一种以海绵为填料的替代性滴滤器。海绵保留的污泥中含有各种各样的生物，包括微型动物。稳定同位素分析揭示了滞留污泥和微动物（水螨和蝇幼虫）碳氮稳定同位素比值（δ13C和δ15N）的时空和生物变化。此外，还发现并分析了成虫蝇和蜘蛛。污泥中的δ13C和δ15N在时间和空间上沿反应器方向相似。同位素特征与处理特性有关，如DHS反应器的低硝化效率。同域蝇幼虫的δ13C和δ15N与水螨幼虫的δ13C和δ15N存在差异，说明不同类群间存在差异。有趣的是，水螨的δ13C和δ15N高于残留污泥，这表明它们在食物网中处于更高的营养地位。此外，蜘蛛的δ13C和δ15N值比成蝇高约1‰~ 3‰。这有力地表明，成年苍蝇和蜘蛛之间存在一种捕食关系。

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引用次数: 0

Using pure oxygen aeration to increase recombinant protein production by an Aspergillus oryzae hyphal dispersion strain 利用纯氧曝气提高米曲霉菌丝分散菌株重组蛋白的产量。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-24 DOI: 10.1016/j.jbiosc.2025.10.011

Satoshi Ishibashi , Shunya Susukida , Kiyoaki Muto , Ken Miyazawa , Silai Zhang , Akira Yoshimi , Eiji Tabata , Keietsu Abe

A high growth rate is essential for increasing protein production efficiency in liquid fermentation of filamentous fungi, such as Aspergillus oryzae. However, the increase in culture viscosity due to fungal growth constrains the overall yield. We have demonstrated that culture viscosity is lower in A. oryzae AGΔ-GAGΔ strains, which are deficient in the cell surface polysaccharides α-1,3-glucan (AG) and galactosaminogalactan (GAG), than in the wild-type (WT) strains. Nevertheless, even in aerated fermentation, an increase in AGΔ-GAGΔ viscosity results in oxygen depletion, which limits fungal growth and enzyme production. In this study, we investigated viscosity dynamics and protein production during high-cell-density fermentation of AGΔ-GAGΔ under pure oxygen aeration. Fed-batch cultivation of the WT and AGΔ-GAGΔ strains, expressing recombinant xylanase (XynF1), was used to compare the effects of air and pure oxygen aeration at the same flow rate. At 60 h, AGΔ-GAGΔ under pure oxygen aeration showed higher cell density (1.2× WT under pure oxygen aeration, 2.1× AGΔ-GAGΔ under air aeration) and XynF1 activity (1.8× WT under pure oxygen aeration, 2.3× AGΔ-GAGΔ under air aeration). Under pure oxygen aeration, AGΔ-GAGΔ showed lower viscosity (0.32×) and mixing time (0.50×) than WT. At 60 h, fine mycelial pellets (micropellets; 200–700 μm) were clearly observed in AGΔ-GAGΔ under pure oxygen but not under air aeration. These findings suggest that oxygen enrichment during AGΔ-GAGΔ cultivation mitigated the increase in viscosity, thereby promoting higher cell density and protein production. The formation of micropellets in AGΔ-GAGΔ likely contributed to a reduction in culture viscosity.

在米曲霉等丝状真菌的液体发酵过程中，提高蛋白质的生产效率需要较高的生长速率。然而，由于真菌生长而增加的培养粘度限制了总体产量。我们已经证明，与野生型（WT）菌株相比，缺乏细胞表面多糖α-1,3-葡聚糖（AG）和半乳糖半乳聚糖（GAG）的米芽孢杆菌AGΔ-GAGΔ菌株的培养粘度较低。然而，即使在曝气发酵中，AGΔ-GAGΔ粘度的增加也会导致氧气消耗，从而限制真菌的生长和酶的产生。在这项研究中，我们研究了在纯氧曝气下AGΔ-GAGΔ高密度发酵过程中的粘度动力学和蛋白质产量。对WT和AGΔ-GAGΔ菌株进行补料分批培养，表达重组木聚糖酶（XynF1），比较相同流速下空气和纯氧曝气的效果。60 h时，纯氧曝气AGΔ-GAGΔ的细胞密度更高（纯氧曝气1.2× WT，空气曝气2.1× AGΔ-GAGΔ）， XynF1活性更高（纯氧曝气1.8× WT，空气曝气2.3× AGΔ-GAGΔ）。在纯氧曝气条件下，AGΔ-GAGΔ的黏度（0.32×）和混合时间（0.50×）均低于WT。60 h时，AGΔ-GAGΔ在纯氧条件下明显可见到细小的菌丝球（微球；200-700 μm），而在空气曝气条件下则没有。这些发现表明，AGΔ-GAGΔ培养过程中的氧富集减轻了黏度的增加，从而促进了更高的细胞密度和蛋白质的产生。AGΔ-GAGΔ中微球的形成可能有助于降低培养粘度。

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引用次数: 0

Selective vanillate production from sugarcane bagasse-derived aromatic compounds using an engineered Pseudomonas sp. NGC7-based strain 利用工程假单胞菌sp. ngc7为基础的菌株从甘蔗渣衍生的芳香化合物中选择性生产香草。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-21 DOI: 10.1016/j.jbiosc.2025.11.001

Md. Jahangir Alam , Naoya Kodama , Kazuma Ikeda , Kanami Muraki , Yudai Higuchi , Masaya Fujita , Naofumi Kamimura , Eiji Masai , Hiroyuki Kurihara , Tomonori Sonoki

In this study, Pseudomonas sp. NGC7 was engineered to selectively produce vanillate (VA) from aromatic compounds in the sugarcane bagasse alkaline extract. A VA O-demethylase gene (vanA4B4)-disrupted strain derived from NGC7 grew and produced VA from the extract containing no saccharides. The organic acids in the extract promoted the strain to grow. The aromatics in the extract were further concentrated by the solid phase extraction with DIAION HP20 resin, and the organic acids were fractionated into the flow-through fraction. A fed-batch culture of NGC7ΔvanA4B4 strain using this concentrated alkaline extract exhibited increased VA production; however, the accumulation of syringate (SA) and 4-hydroxybenzoate (HBA) was also observed along with VA production, despite the strain possessing the genes responsible for SA and HBA degradation. Analysis of the mutants capable of degrading SA while producing VA revealed that mutations in vanR2, a transcriptional repressor of the genes responsible for SA degradation, enabled SA degradation during VA production. In addition, the expression of praI, an HBA hydroxylase derived from Paenibacillus sp. JJ-1b, was suitable for efficient HBA degradation. Thus, the mutation in vanR2 and the expression of praI represented the key engineering strategies for achieving the selective VA production. As the growth of the engineered strains was promoted by the organic acids present in the extract, VA production from the concentrated extract was evaluated in a flow-through-based medium supplemented with mineral salts and metals. Finally, the engineered VA-producing strain produced 4.30 mM VA selectively at a yield of 77 mol% in the practical medium.

本研究对假单胞菌sp. NGC7进行了工程改造，使其能够从甘蔗渣碱性提取物中的芳香化合物中选择性地产生香草酸（VA）。从NGC7衍生的VA o -去甲基化酶基因（vanA4B4）中断菌株生长并从不含糖的提取物中产生VA。提取物中的有机酸促进了菌株的生长。用DIAION HP20树脂固相萃取进一步浓缩提取液中的芳烃，将有机酸分馏成流动馏分。使用该浓缩碱性提取物对NGC7ΔvanA4B4菌株进行分批补料培养，VA产量增加；然而，尽管菌株具有负责SA和HBA降解的基因，但在产生VA的同时，也观察到紫丁香酸（SA）和4-羟基苯甲酸（HBA）的积累。对能够降解SA同时产生VA的突变体的分析表明，vanR2（负责SA降解的基因的转录抑制因子）的突变使SA在VA产生过程中降解。此外，源自Paenibacillus sp. JJ-1b的HBA羟化酶praI的表达适合于HBA的高效降解。因此，vanR2的突变和praI的表达代表了实现选择性VA产生的关键工程策略。由于提取物中存在的有机酸促进了工程菌株的生长，因此在添加无机盐和金属的流动培养基中评估了浓缩提取物的VA产量。最后，该工程产VA菌株在实际培养基中选择性地产生4.30 mM VA，产率为77 mol%。

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引用次数: 0

One-pot synthesis of β-alanine from 1,3-diaminopropane using two-enzyme cascade in cell-free biotransformation 用双酶级联法在无细胞生物转化中一锅法合成1,3-二氨基丙烷β-丙氨酸。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-17 DOI: 10.1016/j.jbiosc.2025.10.013

Shanthi Shanmugasundaram, Anitha Janet Roshni Yesudhas, Andrea Kagoo, Ramalingam Subramanian

A novel two-enzyme cascade one-pot synthesis of β-alanine from 1,3-diaminopropane (DAP) has been developed. In the first step, DAP was oxidized to 3-aminopropionaldehyde (3-APAL) by diamine oxidase (DAO). In the second step, 3-APAL was oxidized to β-alanine by 3-APAL dehydrogenase (APALDH). Catalase and NADH oxidase were employed to degrade the by-product, H₂O₂, and to regenerate the cofactor, NAD⁺. DAO specific to DAP has not been reported in any prokaryote. Therefore, initial proof of concept was established using commercial eukaryotic DAO_pk (from porcine kidney) and catalase (from bovine liver), along with recombinant APALDH and NOX enzymes sourced from Arthrobacter crystallopoietes and Lactococcus lactis, respectively. β-Alanine was successfully produced via this pathway; 12.2 mM (1.1 g/L) was formed in 41 h with 12 % conversion. To increase the reaction rate, DAO with higher specific activity was identified from Arthrobacter pascens (DAO_Ap). The optimum pH and temperature of DAO_Ap were determined to be 9.0 and 37 °C, respectively. Batch enzymatic biotransformation achieved 6 % conversion, yielding 0.33 mM (29 mg/L) β-alanine in 4 h. The low titre in batch conversion was attributed to substrate inhibition affecting DAO_Ap, NOX, and catalase. Fed-batch enzymatic biotransformation was conducted to overcome substrate inhibition, yielding 47 % conversion, with 2.34 mM (63 mg/L) β-alanine formation in 4 h. Approximately a 7.5-fold increase in conversion was achieved using fed-batch enzymatic biotransformation. This study accomplished a novel two-enzyme cascade biotransformation strategy for one-pot β-alanine synthesis from DAP.

以1,3-二氨基丙烷（DAP）为原料，采用双酶级联一锅法合成β-丙氨酸。第一步，二胺氧化酶（DAO）将DAP氧化为3-氨基丙醛（3-APAL）。第二步，3-APAL脱氢酶（APALDH）将3-APAL氧化为β-丙氨酸。过氧化氢酶和NADH氧化酶被用来降解副产物H2O2，并再生辅助因子NAD+。在任何原核生物中未见DAP特异性DAO的报道。因此，最初的概念证明是使用商业真核DAOpk（来自猪肾）和过氧化氢酶（来自牛肝脏），以及分别来自节杆菌晶体和乳酸球菌的重组APALDH和NOX酶建立的。β-丙氨酸通过该途径成功生成；以12%的转化率，在41 h内形成12.2 mM （1.1 g/L）。为了提高反应速率，从pascens节杆菌（Arthrobacter pascens, DAOAp）中鉴定出具有较高比活性的DAO。测定了DAOAp的最佳pH为9.0℃，最佳温度为37℃。批量酶生物转化达到6%的转化率，在4小时内产生0.33 mM (29 mg/L) β-丙氨酸。批量转化的低滴度归因于底物对DAOAp、NOX和过氧化氢酶的抑制作用。为了克服底物抑制，进行了补料批式酶生物转化，转化率为47%，在4小时内生成2.34 mM (63 mg/L) β-丙氨酸。使用补料批式酶生物转化，转化率提高了约7.5倍。本研究完成了一种新的双酶级联生物转化策略，用于一锅法合成β-丙氨酸。

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引用次数: 0

Biodegradation of recalcitrant environmental pollutants by white-rot fungi 白腐菌降解难降解环境污染物的研究。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-17 DOI: 10.1016/j.jbiosc.2025.10.010

Mingdong Chang , Ru Yin , Jianqiao Wang , Nana Wang , Pengfei Xiao

The understanding of white-rot fungi (WRF) and their role in degrading recalcitrant environmental pollutants has significantly advanced due to developments in bioremediation research. Considerable progress has been made in elucidating the degradation capabilities of WRF against lots of environmental pollutants. In this review, research hotspots on the degradation of WRF were identified through bibliometric analysis. Key findings from systematic studies on the degradation of polycyclic aromatic hydrocarbons (PAHs) and bisphenols by WRF are synthesized and discussed. Furthermore, insights into the molecular and genetic basis underlying the enzymatic systems responsible for the degradation of PAHs and bisphenols are highlighted. Advancements and challenges in understanding the degradation capabilities and degradation mechanisms are examined in order to identify opportunities for developing more effective strategies to harness the bioremediation potential of WRF.

由于生物修复研究的发展，人们对白腐菌及其在降解难降解环境污染物中的作用的了解取得了显著进展。WRF对多种环境污染物的降解能力的研究取得了长足的进展。本文通过文献计量学分析，确定了WRF降解的研究热点。综述了WRF降解多环芳烃（PAHs）和双酚类化合物的系统研究成果。此外，深入了解的分子和遗传基础背后的酶系统负责降解多环芳烃和双酚被强调。研究了在理解降解能力和降解机制方面取得的进展和面临的挑战，以确定制定更有效的战略以利用WRF的生物修复潜力的机会。

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Aerobic degradation characteristics of cis-1,2-dichloroethene by Pseudonocardia sp. D17: Degradation kinetics, putative degradation pathways, and involvement of soluble di-iron monooxygenases in the initial oxidation pseudoncardidia sp. D17对顺式-1,2-二氯乙烯的好氧降解特性：降解动力学，假定的降解途径，以及可溶性二铁单加氧酶在初始氧化中的作用

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-12 DOI: 10.1016/j.jbiosc.2025.10.012

Ryugo Nishimine , Yuna Kaneko , Shinpei Fujiwara , Daisuke Inoue , Masahiro Takeo , Michihiko Ike

Pseudonocardia sp. D17 (D17) is a novel strain capable of aerobically metabolizing cis-1,2-dichloroethene (cDCE), a persistent contaminant in soil and groundwater. This study aimed to investigate the cDCE degradation characteristics of D17 with respect to kinetics, associated degradative enzymes, and degradation pathways. Degradation experiments with cDCE concentrations ranging from 0.267 to 91.3 μM revealed that D17 can efficiently degrade cDCE across this range. The maximum specific degradation rate and half saturation constant for cDCE degradation by D17 were estimated to be 0.418 ± 0.045 nmol/mg-protein/min and 38.5 ± 9.2 μM, respectively. Heterologous expression experiments demonstrated that both group 5 soluble di-iron monooxygenases in D17, namely tetrahydrofuran and propane monooxygenases, can catalyze cDCE degradation with higher catalytic activity observed in the former. This suggests their involvement in cDCE degradation by D17. It was also proposed that D17 completely dechlorinates cDCE through multiple pathways to generate glyoxylic acid, which is either mineralized or incorporated into the glyoxylate cycle, with a minor portion being converted to oxalic acid as a dead-end product. These findings provide novel insights into metabolic aerobic cDCE biodegradation and highlight the potential of D17 as a bioremediation agent.

伪心藻sp. D17 （D17）是一种能够代谢土壤和地下水中持久性污染物顺-1,2-二氯乙烯（cDCE）的新型菌株。本研究旨在从动力学、相关降解酶和降解途径等方面探讨D17对cDCE的降解特性。在0.267 ~ 91.3 μM的cDCE浓度范围内，D17能有效降解cDCE。D17降解cDCE的最大特定降解速率和半饱和常数分别为0.418±0.045 nmol/mg-protein/min和38.5±9.2 μM。异源表达实验表明，D17中第5组可溶性二铁单加氧酶，即四氢呋喃单加氧酶和丙烷单加氧酶都能催化cDCE降解，且前者的催化活性更高。这表明它们参与了D17对cDCE的降解。也有人提出，D17通过多种途径使cDCE完全脱氯生成乙醛酸，乙醛酸要么矿化，要么并入乙醛酸循环，一小部分作为死端产物转化为草酸。这些发现为代谢有氧cDCE生物降解提供了新的见解，并突出了D17作为生物修复剂的潜力。

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