Journal of bioscience and bioengineering最新文献_第5页

Isolation and characterization of a novel halotolerant selenate-reducing bacterium, Citrobacter koseri Y2 一种新的耐盐硒酸盐还原菌Citrobacter koseri Y2的分离与鉴定。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-30 DOI: 10.1016/j.jbiosc.2025.10.004

Shunsuke Okahata , Yuya Ueda , Yuki Kikuchi , Takuya Naoe , Daisuke Inoue , Hideo Dohra , Hiroshi Nishikawa , Michihiko Ike

The biological treatment of selenium-containing wastewater has attracted attention as a cost-effective and eco-friendly technology. However, the inhibitory effect of high salinity and oxygen in wastewater on bacterial selenate/selenite-reducing abilities hinders their practical use. In this study, a unique halotolerant facultative anaerobe, Citrobacter koseri Y2, which can reduce selenate under both aerobic and anaerobic conditions, was isolated and characterized, including a whole genome analysis. Strain Y2 reduced 1 mM selenate and selenite, and 0.4 mM selenate and 1 mM selenite to elemental selenium within 4 d at 3 % (w/v) NaCl under aerobic and anaerobic conditions. Regarding the mechanisms underlying selenate reduction, genes for selenate reductases, YnfE and YnfF, and nitrate reductases were identified in the genome of strain Y2. Selenate reduction by strain Y2 was inhibited in the presence of tungstate, confirming the involvement of molybdoenzymes in this process. These results indicate that strain Y2 is a promising bioagent for the treatment of selenium-containing wastewater.

含硒废水的生物处理作为一种经济、环保的技术受到了广泛的关注。然而，废水中高盐度和高氧对细菌硒酸盐/亚硒酸盐还原能力的抑制作用阻碍了它们的实际应用。在这项研究中，分离和表征了一种独特的耐盐兼性厌氧菌Citrobacter koseri Y2，它可以在好氧和厌氧条件下降低硒酸盐，包括全基因组分析。菌株Y2在3% (w/v) NaCl条件下，在4 d内还原1 mM硒酸盐和亚硒酸盐，并将0.4 mM硒酸盐和1 mM亚硒酸盐还原为单质硒。关于硒酸盐还原的机制，在菌株Y2的基因组中发现了硒酸还原酶、YnfE和YnfF以及硝酸盐还原酶的基因。钨酸盐的存在抑制了菌株Y2对硒酸盐的还原，证实了钼酶参与了这一过程。这些结果表明，菌株Y2是一种很有前途的处理含硒废水的生物制剂。

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引用次数: 0

13C-metabolic flux analysis of K562 cells before and after differentiation into erythroid reveals a metabolic shift toward oxidative metabolism K562细胞向红系分化前后的13c代谢通量分析显示其代谢向氧化代谢转变。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-30 DOI: 10.1016/j.jbiosc.2025.10.002

Eisuke Mochizuki , Nobuyuki Okahashi , Takeo Taniguchi , Fumio Matsuda

In regenerative medicine, it is crucial to discover the key factors associated with erythroid differentiation for efficient production of artificial red blood cells. One such factor is erythroid metabolism as erythroid cells dynamically coordinate their metabolic processes to obtain energy for proper differentiation. However, the details of these metabolic changes are not well understood. In this study, we aimed to analyze the metabolism of K562, a cell line that differentiates into erythroid cells using ¹³C-metabolic flux analysis. The results showed that differentiated cells decreased glycolytic flux and increased TCA cycle flux compared with undifferentiated cells, indicating a shift to oxidative metabolism via differentiation. Based on the finding, the inhibition of ATP synthase by oligomycin treatment significantly suppressed differentiation of K562 cells, suggesting that the activation of oxidative metabolism is required for proper differentiation of K562 cells.

在再生医学中，发现与红细胞分化相关的关键因素对于有效地生产人造红细胞至关重要。其中一个因素是红细胞代谢，红细胞动态协调其代谢过程以获得适当分化所需的能量。然而，这些代谢变化的细节尚不清楚。在这项研究中，我们旨在利用13c代谢通量分析K562这一分化为红系细胞的细胞系的代谢。结果表明，与未分化的细胞相比，分化的细胞糖酵解通量降低，TCA循环通量增加，表明细胞通过分化向氧化代谢转变。综上所述，寡霉素处理对ATP合酶的抑制显著抑制了K562细胞的分化，提示K562细胞的正常分化需要激活氧化代谢。

引用次数: 0

Evaluation of the skin whitening efficiency of Staphylococcus epidermidis fermentation broth and its oligopeptides 表皮葡萄球菌发酵液及其寡肽的皮肤美白效果评价。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-29 DOI: 10.1016/j.jbiosc.2025.10.001

Wenlin Geng, Ming Li, Yuhua Cao

In this work, two types of Staphylococcus epidermidis fermentation broth were prepared in beef-protein medium and beef-protein medium with glucose, named as SFB and Glu-SFB. As a positive control, 0.5 mg/mL kojic acid was utilized, which led to a 33.1 ± 1.32 % drop in melanin content and a 30.9 ± 2.95 % reduction in tyrosinase activity in B16–F10 cells. After treatment with SFB and Glu-SFB, the intracellular melanin content diminished by 35.4 ± 0.67 % and 48.5 ± 1.36 %, while tyrosinase activity declined by 59.1 ± 1.49 % and 64.4 ± 2.03 %, respectively. The two S. epidermidis fermentation broth markedly diminish intracellular melanin concentrations and tyrosinase activity, leading to a whitening effect. The whitening efficacy of Glu-SFB surpasses that of SFB and exceeds that of 0.5 mg/mL kojic acid. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with untargeted metabolomics analysis was utilized to identify differential metabolites. Six oligopeptides were identified as Trp-Phe-Tyr-Leu (WFYL), Gln-Ile-Gly-Pro (QIGP), Val-Arg-Phe-Ile (VRFI), Tyr-Ile-Arg (YIR), Glu-Gln-Ile-Trp (EQIW), and His-Gly-Tyr-Lys (HGYK), exhibiting greater relative abundance in Glu-SFB than in SFB. At a dosage of 0.1 mg/mL, oligopeptide exhibits a greater capacity to diminish intracellular melanin levels and tyrosinase activity compared to 0.5 mg/mL kojic acid. Gly-SFB is prepared by replacing glucose with glycerol, the relative concentration of oligopeptides in Gly-SFB is positioned between that of SFB and Glu-SFB, while its whitening efficacy is similarly intermediate between SFB and Glu-SFB. Western blot research shown that all the S. epidermidis fermentation broths may suppress the expression of tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and microphthalmia-associated transcription factor (MITF), which is the primary mechanism underlying the whitening impact of these broths.

本研究在牛肉蛋白培养基和牛肉蛋白加葡萄糖培养基中制备了两种表皮葡萄球菌发酵液，分别命名为SFB和Glu-SFB。以0.5 mg/mL的曲酸作为阳性对照，B16-F10细胞黑色素含量下降33.1±1.32%，酪氨酸酶活性下降30.9±2.95%。经SFB和Glu-SFB处理后，细胞内黑色素含量分别下降35.4%±0.67%和48.5%±1.36%，酪氨酸酶活性分别下降59.1%±1.49%和64.4±2.03%。两种表皮葡萄球菌发酵液显著降低细胞内黑色素浓度和酪氨酸酶活性，从而达到美白效果。Glu-SFB的美白效果超过SFB，超过0.5 mg/mL的曲酸。采用液相色谱-串联质谱（LC-MS/MS）结合非靶向代谢组学分析鉴定差异代谢物。6个寡肽分别为：trp - fe - il - gly - pro （QIGP）、val - arg - fe - ile （VRFI）、tyr - il - arg （YIR）、glu - gln - il - trp （EQIW）和his - gln - il - lys (HGYK)，其中Glu-SFB中的相对丰度高于SFB。与0.5 mg/mL的曲酸相比，在0.1 mg/mL的剂量下，寡肽表现出更大的降低细胞内黑色素水平和酪氨酸酶活性的能力。Gly-SFB是用甘油代替葡萄糖制备的，Gly-SFB中寡肽的相对浓度介于SFB和Glu-SFB之间，其美白功效也同样介于SFB和Glu-SFB之间。Western blot研究表明，所有表皮葡萄球菌发酵液均可抑制酪氨酸酶（TYR）、酪氨酸酶相关蛋白-1 （TRP-1）、酪氨酸酶相关蛋白-2 （TRP-2）和小眼相关转录因子（MITF）的表达，这是这些发酵液美白作用的主要机制。

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引用次数: 0

Conversion of arachidonic acid into 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid by Torula dematia NBRC 6213 花生四烯酸转化为14,15,19-三羟基二糖-5,8,11-三烯酸的研究。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-07 DOI: 10.1016/j.jbiosc.2025.09.004

Wataru Fujii , Michiki Takeuchi , Si-Bum Park , Kazuki Yagi , Shunta Nakamura , Ei-Tora Yamamura , Jun Ogawa

Lipids are one of the three major nutrients that play important roles as sources of energy and as components of cell membranes. Fatty acid metabolites exhibit physiological activities. Among fatty acid metabolites, some hydroxy arachidonic acids (ARAs) have bioactive functions. In this study, we found that Torula dematia NBRC 6213 converts ARA into unknown fatty acids. Liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry, and nuclear magnetic resonance (NMR) analyses were conducted to identify this unknown fatty acid. The unknown fatty acid was identified as 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid. Its production was maximized when T. dematia was cultivated in potato dextrose broth (PDB) medium and reacted for 12 h at pH 7.0 and 28 °C. In addition, multiple intermediates were formed during the conversion of ARA to 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid. LC-MS analysis revealed molecular weights of 320, 336, and 338. This suggests that ARA conversion occurs via the hydroxylation, epoxidation, and hydrolysis of the epoxy group. T. dematia also converts other unsaturated fatty acids into similarly oxidized fatty acids.

脂质是三种主要营养素之一，作为能量来源和细胞膜成分起着重要作用。脂肪酸代谢产物具有生理活性。在脂肪酸代谢产物中，一些羟基花生四烯酸具有生物活性。在这项研究中，我们发现Torula dematia NBRC 6213将ARA转化为未知的脂肪酸。采用液相色谱-质谱（LC-MS）、气相色谱-质谱（gc - ms）和核磁共振（NMR）分析对该未知脂肪酸进行鉴定。未知脂肪酸鉴定为14,15,19-三羟基二糖-5,8,11-三烯酸。在马铃薯葡萄糖肉汤（PDB）培养基中培养，在pH 7.0、28℃条件下反应12 h，其产量最大。此外，ARA在转化为14,15,19-三羟基二糖-5,8,11-三烯酸的过程中形成了多个中间体。LC-MS分析显示分子量为320、336和338。这表明ARA转化通过羟基化、环氧化和环氧基水解发生。脱脂腺也将其他不饱和脂肪酸转化为类似的氧化脂肪酸。

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引用次数: 0

Characterization of a molting-related chitinase from a land crab, Chiromantes haematocheir 一种陆地蟹（Chiromantes haematocheir）几丁质酶的特性。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-07 DOI: 10.1016/j.jbiosc.2025.09.006

Yuma Nagakura, Katsuhide Miyake

Chitinases play an important role in many biological processes, including molting, digestion, and immunity in crustaceans. This study represents an attempt to apply chitinases in the field of biotechnology, detecting chitinase mRNAs from a land crab, Chiromantes haematocheir, via transcriptome analysis and analyzing their properties. Seven chitinase genes were detected from the RNA-seq data of midgut glands. Among these genes, TRINITY_DN29294 transcripts accounted for virtually all of total expression of the chitinases. The 29294 cDNA contained a 1467 bp open reading frame, coded for 488 amino acid residues, and was classified into the GH18 chitolectin chitotriosidase and the group 3 crab chitinase. The expression of the 29294-chitinase mRNA was detected in all tissues, with the highest levels expressed in the midgut glands. The transcripts increased significantly in the early post-molted crab compared to the non-molting crab. These results suggest that 29294-chitinase plays important roles in the molting process. While the recombinant 29294-chitinase over-produced in Escherichia coli did not show any activity, the enzyme expressed in Pichia pastoris exhibited sufficient activity. The 29294-chitinase had its optimal pH 3.0. The optimal temperature was relatively high at 45 °C. The enzyme hydrolyzed both soluble and crystalline substrates.

几丁质酶在甲壳类动物的蜕皮、消化和免疫等生物过程中起着重要作用。本研究尝试将几丁质酶应用于生物技术领域，通过转录组分析检测陆地蟹Chiromantes haematocheir的几丁质酶mrna并分析其性质。从中肠腺的RNA-seq数据中检测到7个几丁质酶基因。在这些基因中，TRINITY_DN29294转录本几乎占几丁质酶总表达量的全部。29294 cDNA包含1467 bp的开放阅读框，编码488个氨基酸残基，被分类为GH18壳聚糖壳三醇苷酶和第3组螃蟹几丁质酶。29294-几丁质酶mRNA在各组织中均有表达，其中以中肠腺表达量最高。与未蜕皮的蟹相比，早期蜕皮蟹的转录本显著增加。上述结果表明，29294-几丁质酶在玉米的蜕皮过程中起着重要作用。在大肠杆菌中过量产生的重组29294-几丁质酶没有表现出任何活性，而在毕赤酵母中表达的酶表现出足够的活性。29294-几丁质酶的最佳pH值为3.0。最佳温度为45℃。这种酶既能水解可溶性底物，也能水解结晶底物。

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引用次数: 0

Design of bispecific antibody Fc region employing a shark-human chimeric and asymmetric format 采用鲨鱼-人嵌合和不对称格式设计双特异性抗体Fc区。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-01 DOI: 10.1016/j.jbiosc.2025.09.001

Ryota Munetomo , Aiko Inoue , Masayoshi Onitsuka

Bispecific antibodies (BsAbs) can bind to two antigens simultaneously and have undergone rapid advancements in recent years owing to their ability to enable novel mechanisms of action that are unachievable using conventional monoclonal antibodies (mAbs). However, the structural complexity of BsAbs remains a problem during product development. One of these problems is the presence of impurities and by-products. Although BsAbs with the human Fc region must be assembled using heterogeneous polypeptide chains, undesired by-products from unpaired and mispaired chain components can contaminate them. These by-products are difficult to remove in the purification process because their physicochemical properties resemble those of the target BsAb with correct pairing. Here, we designed a novel Fc region for enhanced BsAbs in which the human CH2 domain on one side of the Fc region was replaced with the C2 domain from an immunoglobulin new antigen receptor (IgNAR) shark antibody. The designed BsAbs with chimeric and asymmetric Fcs exhibited separate pH elution profiles against soluble aggregates in protein A affinity chromatography. An overlapping elution profile corresponding to the by-product homogeneous chain observed in human Fc BsAbs was not detected in shark C2-introduced BsAbs. Although another homogeneous by-product was observed in the designed BsAb, introducing N-glycosylation at C2 significantly improved this problem. Additionally, BsAbs with the designed Fc demonstrated higher stability in both the colloidal and structural aspects. This study is the first approach for the chimeric and asymmetric design of Fc using a shark-derived constant domain and offers a novel alternative for BsAb development.

双特异性抗体（BsAbs）可以同时结合两种抗原，近年来取得了快速进展，因为它们能够实现传统单克隆抗体（mab）无法实现的新作用机制。然而，在产品开发过程中，bsab结构的复杂性仍然是一个问题。其中一个问题是杂质和副产品的存在。虽然具有人类Fc区的bsab必须使用异质多肽链进行组装，但来自未配对和错配链成分的不希望的副产物会污染它们。这些副产物在纯化过程中很难去除，因为它们的物理化学性质与正确配对的目标BsAb相似。在这里，我们设计了一个新的Fc区域用于增强bsab，其中Fc区域一侧的人CH2结构域被免疫球蛋白新抗原受体（IgNAR）鲨鱼抗体的C2结构域取代。在蛋白A亲和层析中，嵌合Fcs和不对称Fcs的bsab对可溶性聚集体表现出不同的pH洗脱谱。与在人类Fc bsab中观察到的副产物均相链相对应的重叠洗脱谱在鲨鱼c2引入的bsab中未检测到。虽然在设计的BsAb中观察到另一个均匀的副产物，但在C2处引入n -糖基化显着改善了这个问题。此外，具有设计Fc的bsab在胶体和结构方面都表现出更高的稳定性。该研究首次采用鲨鱼衍生的恒定结构域进行Fc嵌合和非对称设计，为BsAb的开发提供了一种新的选择。

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引用次数: 0

Development of a method for quantifying metabolites in Escherichia coli colonies using hyperspectral imaging 利用高光谱成像技术定量大肠杆菌菌落代谢物方法的建立。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-01 DOI: 10.1016/j.jbiosc.2025.09.005

Manami Takama , Takatoshi Suematsu , Takayuki Okano , Shumpei Asamizu , Takahiro Bamba , Tomohisa Hasunuma

Fermentation by microorganisms has attracted attention for the synthesis of bulk and fine chemicals with high added value, including pharmaceutical intermediates. To accelerate the development of high-producing microbial strains, a rapid screening method is warranted. This study aimed to develop a novel, nondestructive approach to quantify metabolite production in microbial colonies using hyperspectral imaging (HSI). As a model, we examined the heterologous production of 1,3,5-trihydroxyanthraquinone (AQ256), an anthraquinone with antimicrobial and anticancer activities, using Escherichia coli. Fluorescence spectral data from HSI, along with AQ256 concentrations measured via high-performance liquid chromatography, were used to construct regression models. In addition, red-green-blue (RGB)-based models were developed, as AQ256 exhibits a characteristic reddish-brown color. Four regression models were compared: multiple linear regression, partial least squares regression (PLSR), support vector regression, and random forest regression. Among them, the PLSR model based on HSI data showed the highest prediction accuracy (R² = 0.75 ± 0.23, root mean square error = 0.08 ± 0.02, mean absolute error = 0.07 ± 0.02). In particular, it outperformed the RGB-based model in extrapolation beyond the training data. These findings demonstrate that the HSI-based method enables accurate, nondestructive quantification of metabolites and has strong potential for high-throughput screening of microbial strains that produce various valuable compounds at elevated yields.

微生物发酵已成为合成高附加值原料药和精细化学品（包括医药中间体）的研究热点。为了加速高产微生物菌株的开发，需要一种快速筛选方法。本研究旨在开发一种新的、无损的方法，利用高光谱成像（HSI）来量化微生物菌落中代谢物的产生。作为模型，我们研究了利用大肠杆菌外源生产具有抗菌和抗癌活性的蒽醌类化合物1,3,5-三羟基蒽醌（AQ256）。HSI的荧光光谱数据与高效液相色谱测定的AQ256浓度一起构建回归模型。此外，基于红-绿-蓝（RGB）的模型被开发出来，因为AQ256表现出红棕色的特征。比较了多元线性回归、偏最小二乘回归、支持向量回归和随机森林回归四种回归模型。其中，基于HSI数据的PLSR模型预测精度最高（R2 = 0.75±0.23，均方根误差= 0.08±0.02，平均绝对误差= 0.07±0.02）。特别是，它在训练数据之外的外推方面优于基于rgb的模型。这些发现表明，基于si的方法能够准确、无损地定量代谢物，并具有高通量筛选微生物菌株的强大潜力，这些微生物菌株能以高产量产生各种有价值的化合物。

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引用次数: 0

Importance rapid initial decline in oxidation–reduction potential, followed by an increase in extracellular electron transport activities, for the rapid onset of indigo reduction 重要的是氧化还原电位的快速初始下降，随后是细胞外电子传递活动的增加，对靛蓝还原的快速开始。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-09-29 DOI: 10.1016/j.jbiosc.2025.08.008

Nowshin Farjana , Hiromitsu Furukawa , Kensuke Igarashi , Souichiro Kato , Isao Yumoto

In most complex microbial systems, the ideal process underlying transitional microbial changes that lead to the formation of functional states is not fully elucidated. To understand the basis for the occurrence of indigo reduction, we analyzed the prerequisites causing transitional shifts in microflora that lead to the indigo-reducing state. To this end, timing of wheat bran (WB) addition, during indigo fermentation process using sukumo (composted leaves of Polygonum tinctorium L.) as the inoculum, substrate, and indigo source, were varied. Early initiation of indigo reduction was achieved through the early proliferation of obligate anaerobic Alkalicella caledoniensis followed by Alkalibacterium spp. or Evansella vedderi. Although it can be predicted that Alkalicella caledoniensis exhibits extracellular electron transport (EET) activity, to promote even effective reduction of indigo, Alkalibacterium spp. or E. vedderi, which have the EET gene sequence series and exert strong metabolic abilities, should emerge using WB. The emergence of Alkalicella caledoniensis was associated with drastic a decrease in bacterial diversity and a concurrent rapid decline in oxidation–reduction potential (ORP). The rate and extent of Alkalicella caledoniensis appearance depended on the rate of ORP reduction. Multivariate analysis (i.e., RDA) revealed that Alkalicella caledoniensis directed the initial drastic changes of microbiota, aligning with the decline in ORP. Prior to these major microbial shifts oxygen consumption by aerobic bacteria utilizing sukumo initiated the ORP decrease. These findings contribute to understanding the approach to steer the initially highly diverse bacterial community during early fermentation toward rapid induction of indigo reduction.

在大多数复杂的微生物系统中，导致功能状态形成的过渡性微生物变化的理想过程尚未完全阐明。为了了解靛蓝还原发生的基础，我们分析了导致靛蓝还原状态的微生物区系过渡变化的先决条件。为此，在以黄蓼（Polygonum tinctorium L.）堆肥叶为接种物、底物和靛蓝源的靛蓝发酵过程中，小麦麸皮（WB）的添加时间不同。靛蓝还原的早期开始是通过专性厌氧caledoniensis的早期增殖实现的，其次是Alkalibacterium sp .或Evansella vedderi。虽然可以预测caledoniensis具有胞外电子传递（extracellular electron transport， EET）活性，但为了促进靛蓝的有效还原，还需要利用WB来研究具有EET基因序列序列且具有较强代谢能力的Alkalibacterium spp.或e.w edderi。caledoniensis的出现伴随着细菌多样性的急剧减少和氧化还原电位（ORP）的快速下降。喀里多碱菌出现的速度和程度取决于ORP还原的速度。多变量分析（即RDA）表明，caledoniensis引导了微生物群最初的剧烈变化，与ORP的下降一致。在这些主要的微生物转变之前，好氧细菌利用sukumo的耗氧量引发了ORP的降低。这些发现有助于理解在早期发酵过程中引导最初高度多样化的细菌群落快速诱导靛蓝还原的方法。

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引用次数: 0

Enrichment of soy protein-derived peptides that decrease pancreatic lipase activity using heat-treated porous silica gel and their relationship with bile acid binding activity 利用热处理多孔硅胶富集降低胰脂肪酶活性的大豆蛋白衍生肽及其与胆汁酸结合活性的关系。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-09-29 DOI: 10.1016/j.jbiosc.2025.09.002

Yusuke Ishii, Yuta Matsunaga, Hirokazu Akiyama, Kazunori Shimizu, Hiroyuki Honda

Excessive lipid absorption is a key factor in obesity. Lipids are solubilized in the gut via bile acid (BA) micelles, where pancreatic lipase hydrolyzes them for absorption. This study aimed to enrich pancreatic lipase inhibitory (PLI) peptides from food protein hydrolysates and clarify their inhibition mechanisms. We used heat-treated porous silica gel (HTSG) to selectively enrich basic and hydrophobic peptides through adsorption–desorption. While HTSG has previously enriched PLI peptides, the mechanism remained unclear. Since basic and hydrophobic peptides can bind strongly to BAs like taurocholic acid, we explored their BA-binding and PLI activities. Pepsin hydrolysates from casein, soybean, pea, and rice endosperm were tested with 1 mM sodium taurocholate (TCA). TCA increased lipase activity over 2.5-fold. Soybean pepsin hydrolysate (SPH) showed notable PLI activity, further enhanced approxiamtely 3-fold after HTSG treatment (SPH (after)). LC–MS/MS of SPH (after) identified 1461 peptides. Among 38 high-abundance peptides (Z ≥ 2) chemically synthesized, 9 inhibited pancreatic lipase in the presence of TCA. BA-binding activity was assessed via micelle disruption. Seven of the nine peptides disrupted over 50 % of micelles. Docking simulation was conducted and peptides that exhibited PLI activity even without TCA and showed TCA-binding activity were predicted to bind directly to pancreatic lipase. In summary, we identified 9 PLI peptides from SPH, most of which inhibit pancreatic lipase by binding to BAs. HTSG-based enrichment offers a promising strategy to obtain bioactive peptides that may serve as functional ingredients for obesity prevention.

脂质吸收过多是肥胖的一个关键因素。脂质通过胆汁酸（BA）胶束在肠道中溶解，胰脂肪酶将其水解以供吸收。本研究旨在从食物蛋白水解物中富集胰脂肪酶抑制肽，并阐明其抑制机制。我们使用热处理多孔硅胶（HTSG）通过吸附-解吸选择性富集碱性肽和疏水性肽。虽然HTSG先前富集了PLI肽，但其机制尚不清楚。由于碱性肽和疏水性肽可以与牛磺酸等ba强结合，我们研究了它们的ba结合和PLI活性。用1mm牛磺胆酸钠（TCA）检测酪蛋白、大豆、豌豆和水稻胚乳的胃蛋白酶水解物。TCA使脂肪酶活性增加了2.5倍以上。大豆胃蛋白酶水解物（SPH）表现出显著的PLI活性，经HTSG处理后，SPH的PLI活性进一步提高了约3倍。SPH的LC-MS/MS鉴定出1461个多肽。在化学合成的38个高丰度肽（Z≥2）中，9个在TCA存在下抑制胰腺脂肪酶。通过胶束破坏来评估ba的结合活性。9个肽中的7个破坏了超过50%的胶束。对接模拟表明，即使没有TCA也具有PLI活性并显示TCA结合活性的肽可以直接与胰脂肪酶结合。总之，我们从SPH中鉴定出9个PLI肽，其中大多数通过与BAs结合来抑制胰脂肪酶。基于htsg的富集为获得生物活性肽提供了一种有前途的策略，这些活性肽可能作为预防肥胖的功能成分。

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引用次数: 0

Whole-genome draft assemblies of Paracoccus pantotrophus DSM 11073 and Paracoccus sp. AS002: Phylogenetics entails classification as Paracoccus versutus AS002 泛养副球菌DSM 11073和副球菌sp. AS002的全基因组草图组装：系统发育需要分类为反副球菌AS002。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-09-27 DOI: 10.1016/j.jbiosc.2025.09.003

Upasana Pal , Denise Bachmann , Linda Fenske , Lars Mathias Blank , Till Tiso

The Gram-stain-negative bacterium Paracoccus spp., from the class Alphaproteobacteria and family Rhodobacteraceae, exhibits exceptional metabolic flexibility, diverse substrate utilization, and tolerance to abiotic stressors. To broaden the biotechnological applications of the genus, comprehensive sequencing, phylogenetic, and physiological characterization were performed between two strains of the genus, Paracoccus pantotrophus DSM 11073 and Paracoccus sp. AS002. Illumina sequencing yielded a total genome size of 4.2 Mbp for P. pantotrophus DSM 11073 and 4.8 Mbp for Paracoccus sp. AS002. Through phylogenetic analysis using the EDGAR software, Paracoccus sp. AS002 shared with Paracoccus versutus DSM 582 the same clade in the phylogenetic tree and an ANI score of 98.9 % indicating that Paracoccus sp. AS002 could be reclassified as P. versutus AS002. The study was extended to compare various attributes of the sequenced genomes and highlight the metabolic versatility of the genus Paracoccus. The use of a wide substrate panel demonstrated the metabolic versatility of the strains, including the PET monomer ethylene glycol, the C1 carbon source formic acid, and renewable carbon sources such as ethanol. Additionally, the ability of the strains to produce bioplastic was assessed, with P. pantotrophus DSM 11073 producing 36 % and P. versutus AS002 28 % polyhydroxybutyrate (% cell dry weight) on glucose, and 40 % and 16 % on 60 mM ethylene glycol, respectively. This study demonstrates the value of sequencing bacterial strains for biotechnological applications and highlights EDGAR's effectiveness in phylogenetic analysis, paving the way for using Paracoccus as a microbial chassis in sustainable biotechnological processes supporting the circular bioeconomy.

革兰氏染色阴性的副球菌属，属于α变形菌纲和红杆菌科，表现出特殊的代谢灵活性，多种底物利用和对非生物应激源的耐受性。为了扩大该属的生物技术应用，对两株pantotrophus Paracoccus DSM 11073和Paracoccus sp. AS002进行了全面的测序、系统发育和生理特性研究。Illumina测序结果显示，P. pantotrophus DSM 11073的总基因组大小为4.2 Mbp，副球菌sp. AS002的总基因组大小为4.8 Mbp。通过EDGAR软件进行系统发育分析，发现副球菌sp. AS002与反苏副球菌DSM 582在系统发育树上具有相同的进化支，ANI评分为98.9%，表明副球菌sp. AS002可以重新归类为反苏副球菌AS002。该研究扩展到比较测序基因组的各种属性，并突出副球菌属的代谢多样性。广泛底物面板的使用证明了菌株代谢的多功能性，包括PET单体乙二醇、C1碳源甲酸和可再生碳源如乙醇。此外，对菌株产生生物塑料的能力进行了评估，P. pantotrophus DSM 11073在葡萄糖上产生36%的聚羟基丁酸盐，P. versutus AS002在60 mM乙二醇上产生28%的聚羟基丁酸盐（细胞干重的%），分别为40%和16%。该研究证明了细菌菌株测序在生物技术应用中的价值，并强调了EDGAR在系统发育分析中的有效性，为利用副球菌作为支持循环生物经济的可持续生物技术过程中的微生物基础铺平了道路。

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