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Multicopy Expression of the Marine Antimicrobial Peptide Spgillcin177-189 in Pichia pastoris for High-Yield Production and Potent Activity Against Foodborne Pathogens. 海洋抗菌肽Spgillcin177-189在毕赤酵母中高产表达及抗食源性致病菌活性研究
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-15 DOI: 10.1007/s12602-025-10874-y
Xianxian Dong, Huiliang Liao, Chang Zhang, Fangyi Chen, Hui Peng, Xiao Hong, Hua Hao, Ming Xiong, Jiahao Ma, Ke-Jian Wang

Bacterial foodborne contamination poses a dual challenge of chemical preservative risks and antibiotic resistance, drives the need for green production of natural antimicrobial alternatives. The reported cationic antimicrobial peptide (AMP) Spgillcin177-189 derived from the Scylla paramamosain, has strong antimicrobial activity against Staphylococcus aureus and clinical isolation strains. To meet industry demand in future, large-scale production of Spgillcin177- 189 is essential. In the study, Pichia pastoris expression system was established for production of the recombinant Spgillcin177- 189 (rSpgillcin177-189). Then, multicopy strategy was selectively designed by employing the Golden Gate assembly technology to efficiently construct multi-copy plasmids, which significantly enhanced the expression level of Spgillcin177- 189. A yield of 126.1 mg/L was harvested with 2.75-fold higher that of the single-copy strain. In addition, the recombinant Spgillcin177 - 189 exhibited potent antibacterial activity against multiple foodborne pathogens within a MIC range of 5.25-84 µg/mL. It also showed effective bactericidal activity and anti-biofilm activity against Staphylococcus aureus and Vibrio parahaemolyticus. rSpgillcin177 - 189 exhibited good thermostability, with no obvious cytotoxicity and hemolytic activity. rSpgillcin177 - 189 may interact with microbial surface components via hydrogen bonding, which were vital for peptide activity in combating bacteria. The rSpgillcin177 - 189 specifically targeting the cell membrane, disrupted bacterial membrane integrity and leading to cell death. This study provided a very feasible genetic engineering strategy for large-scale production of rSpgillcin177 - 189, which will be applied at a lower cost in agricultural and food industries in future.

细菌性食源性污染带来了化学防腐剂风险和抗生素耐药性的双重挑战,推动了对天然抗菌替代品绿色生产的需求。从Scylla paramamosain中分离得到的阳离子抗菌肽Spgillcin177-189对金黄色葡萄球菌和临床分离菌株具有较强的抗菌活性。为了满足未来的工业需求,Spgillcin177- 189的大规模生产是必不可少的。本研究建立了重组Spgillcin177-189 (rSpgillcin177-189)的毕赤酵母表达体系。然后,利用金门组装技术选择性设计多拷贝策略,高效构建多拷贝质粒,显著提高Spgillcin177- 189的表达水平。产量为126.1 mg/L,是单拷贝菌株的2.75倍。此外,重组菌Spgillcin177 - 189对多种食源性致病菌的MIC范围为5.25 ~ 84µg/mL,具有较强的抑菌活性。对金黄色葡萄球菌和副溶血性弧菌也有较好的杀菌和抗生物膜活性。rSpgillcin177 - 189具有良好的热稳定性,无明显的细胞毒性和溶血活性。rSpgillcin177 - 189可能通过氢键与微生物表面组分相互作用,这对抗菌肽的活性至关重要。rSpgillcin177 - 189特异性靶向细胞膜,破坏细菌膜完整性,导致细胞死亡。本研究为rSpgillcin177 - 189的大规模生产提供了一种非常可行的基因工程策略,将在未来以较低的成本应用于农业和食品工业。
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引用次数: 0
Vegetable Juice Fermented with Lactiplantibacillus plantarum WB3813 and Lactiplantibacillus plantarum WB3814 Alleviates H2O2-induced Oxidative Stress in Respiratory Models. 植物乳酸杆菌WB3813和植物乳酸杆菌WB3814发酵的蔬菜汁减轻h2o2诱导的呼吸模型氧化应激
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-12 DOI: 10.1007/s12602-025-10861-3
Ji-Yeon Baek, Seo-Bin Kim, Young-Seo Park, Dae-Kyung Kang, Na-Kyoung Lee, Hyun-Dong Paik

This study investigated the oxidative stress alleviating effects of vegetable juice (VJ) fermented with Lactiplantibacillus plantarum WB3813 and L. plantarum WB3814 using a H2O2-induced A549 cell model. Both WB3813 and WB3814 strains exhibited stable fermentation characteristics in VJ, supporting their suitability as starter cultures for developing plant-based probiotic products. Fermentation enhanced antioxidant activity, with WB3814-VJ showing the highest total polyphenol content (TPC) (204 mg GAE/mL), while WB3813-VJ exhibited the strongest 2,2-diphenyl-1-picrylhydrazyl (DPPH) (38%) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities (68%). Treatment with fermented VJ with WB3813 and WB3814 showed greater cytoprotective effects than LGG in H2O2-induced A549 cells and significantly reduced reactive oxygen species (ROS) production (89.74%-91.67%). Additionally, the expressions of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 were downregulated, while that of antioxidant-related genes, including Nrf2, HO-1, and NQO1, were upregulated. These anti-inflammatory and antioxidant effects were closely associated with the modulation of the NF-κB and Keap1/Nrf2/HO-1 signaling pathways. These findings suggest that VJ fermented with L. plantarum WB3813 and WB3814 can mitigate oxidative stress through its anti-inflammatory and antioxidant activities, supporting its potential as a functional food for respiratory health.

采用h2o2诱导的A549细胞模型,研究了植物乳杆菌WB3813和植物乳杆菌WB3814发酵的蔬菜汁(VJ)对氧化应激的缓解作用。WB3813和WB3814菌株在VJ中表现出稳定的发酵特性,适合作为植物性益生菌产品的发酵剂。发酵增强了其抗氧化活性,其中WB3814-VJ的总多酚含量(TPC)最高(204 mg GAE/mL),而WB3813-VJ的2,2-二苯基-1-picrylhydrazyl (DPPH)(38%)和2,2'-氮基-双(3-乙基苯并噻唑-6-磺酸)(ABTS)自由基清除能力最强(68%)。WB3813和WB3814发酵VJ对h2o2诱导的A549细胞的细胞保护作用优于LGG,显著降低活性氧(ROS)的产生(89.74% ~ 91.67%)。促炎因子TNF-α、IL-1β、IL-6表达下调,抗氧化相关基因Nrf2、HO-1、NQO1表达上调。这些抗炎和抗氧化作用与NF-κB和Keap1/Nrf2/HO-1信号通路的调节密切相关。综上所述,植物乳杆菌WB3813和WB3814发酵的VJ可通过其抗炎和抗氧化活性减轻氧化应激,支持其作为呼吸系统健康功能性食品的潜力。
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引用次数: 0
Assessment of a Novel Synbitotic of Pomegranate Peels Extract and Multi-Specie Probiotics as an Antibiotics Alternative in Ruminants. 石榴皮提取物与多种益生菌合成物作为反刍动物抗生素替代品的研究。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-11 DOI: 10.1007/s12602-025-10822-w
Nesrein M Hashem, Nourhan S Hosny, Nagwa El-Desoky, Gomaa Abo El Lail, Yosra A Soltan, Ahmed A Elolimy, El-Sayed M Abu-Tor, Sobhy M A Sallam

A synbiotic composed of pomegranate peels extract, Saccharomyces (S.) cerevisiae, Lactococcus (L.) lactis, Lactobacillus (Lact) paracasei, and Lact. plantarum was fabricated in free (FPLYS) and (NPLYS) nano-encapsulated forms. The encapsulation efficiency, physichochemical properties, and antioxidant and antimicrobial activities of FPLYS and NPLYS were tested. For in vitro rumen fermentation study, a basal diet was supplemented with FPLYS-H (109 CFU probiotics/g diet), NPLYS-H (109 CFU probiotics/g diet), NPLYS-L (107 CFU probiotics/g diet), 40 mg/kg diet monensin (MONS), or not supplemented (CON). The encapsulation efficiency of gum arabic-gelatin complex for prebiotic and probiotics was 75.26% and 91.80%, respectively. The size and zeta potential of gum arabic-gelatin were 27.72 nm and 11 mV, respectively. NPLYS showed two-time higher antioxidant activity than FPLYS. NPLYS showed antimicrobial activity against Escherichia (E.) coli and Staphylococcus (Staph.) aureus similar to gentamicin, while FPLYS showed lower activity (p < 0.05). NPLYS and FPLYS showed antimicrobial activity against Pseudomonas (Ps.) aeruginosa and antifungal activity against Aspergillus (A.) niger and A. flavus, but lower than getntamicin and fluconazol, respectively (p < 0.05). NPLYS-L significantly increased total gas production compared to CON, MONS, and NPLYS-H. Compared to CON, both MONS and FPLYS-H significantly reduced methane production, followed by NPLYS-H and NPLYS-L. Among treatments, MONS significantly increased propionate, while reducing butyrate and acetate/propionate ration. All treatments increased cellulose digestibility and protozoal count (p < 0.05). NPLYS-L significantly decreased rumen ammonia concentration compared to CON, however other treatments recorded intermediate values. The synbiotic developed in this study can be either used as an antimicrobial agent and/or a rumen fermentation modifier, replacing synthetic antibiotics used for these purposes. Nanoencapsulation process improved biological activities of the synbiotic.

一种由石榴皮提取物、酿酒酵母菌、乳酸乳球菌、副乳酸乳杆菌和乳酸乳杆菌组成的合成物。制备了游离(FPLYS)和(NPLYS)纳米封装形式的植物体。对FPLYS和NPLYS的包封率、理化性质、抗氧化和抗菌活性进行了测试。在体外瘤胃发酵试验中,在基础饲粮中添加FPLYS-H (109 CFU益生菌/g饲粮)、NPLYS-H (109 CFU益生菌/g饲粮)、NPLYS-L (107 CFU益生菌/g饲粮)、40 mg/kg饲粮莫能菌素(MONS)或不添加(CON)。阿拉伯胶-明胶配合物对益生元和益生菌的包封率分别为75.26%和91.80%。阿拉伯胶-明胶的尺寸为27.72 nm, zeta电位为11 mV。NPLYS的抗氧化活性比FPLYS高2倍。NPLYS对大肠杆菌和金黄色葡萄球菌的抑菌活性与庆大霉素相似,而FPLYS对大肠杆菌和金黄色葡萄球菌的抑菌活性较低(p
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引用次数: 0
Compound Lactic Acid Bacteria Alleviate Metabolic Dysregulation in Rhesus Macaques with Chronic Restraint Stress Via Remodeling the Gut Microbiota. 复合乳酸菌通过重塑肠道菌群缓解慢性约束应激恒河猴代谢失调。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-11 DOI: 10.1007/s12602-025-10881-z
Yan Shu, Xiangli Bai, Kaixin Song, Xianguo Li, Chenxing Jian, Yongping Shi, Xiaogang Shu, Si Jin

Chronic stress is a critical risk factor for depression, and prolonged exposure to stressors increases the risk of metabolic dysregulation. Gut microbiota (GM) intervention is a promising therapeutic target for stress-related emotional and behavioral disorders. In the present study, we apply compound lactic acid bacteria (CLAB) (Lacticaseibacillus rhamnosus GG and Enterococcus faecium) administration therapy to investigate the effect of gut microbiota (GM) on chronic restraint stress (CRS)-depression and its related glucose-lipid metabolism disturbance. We established a non-human primate model of CRS depression to mimic the depressive state in humans and applied CLAB for gastric gavage treatment. CRS macaques exhibit marked depressive-like behavior and disrupted GM structure, decreased levels of C1q/tumor necrosis factor-related protein 9 (CTRP9) and increased levels of CTRP5, and induced insulin resistance (IR) and lipid metabolism disorders. Treatment with CLAB normalized GM structure in CRS macaques, increased the abundance of Lactobacillus murinus and Lactobacillus reuteri, and reduced depressive-like behavior in CRS macaques, with a rebound in CTRP9 and HDL-C levels, reduced CTRP5 and TG levels and improved IR. In addition, CLAB may serve as a promising non-pharmacological intervention that alleviates depression-related metabolic dysregulation by modulating the gut microbiota and improving lipid and glucose metabolism.

慢性压力是抑郁症的关键风险因素,长期暴露于压力源会增加代谢失调的风险。肠道菌群(GM)干预是治疗压力相关情绪和行为障碍的一个有希望的治疗靶点。本研究采用复合乳酸菌(lactoaseibacillus rhamnosus GG和Enterococcus faecium)给药治疗,探讨肠道菌群(GM)对慢性抑制性应激(CRS)抑制及其相关的糖脂代谢紊乱的影响。我们建立非人类灵长类动物CRS抑郁模型,模拟人类抑郁状态,并应用CLAB进行灌胃治疗。CRS猕猴表现出明显的抑郁样行为和GM结构破坏,C1q/肿瘤坏死因子相关蛋白9 (CTRP9)水平下降和CTRP5水平升高,并诱导胰岛素抵抗(IR)和脂质代谢紊乱。CLAB标准化GM结构治疗CRS猕猴,增加了鼠乳杆菌和罗伊氏乳杆菌的丰度,减少了CRS猕猴的抑郁样行为,CTRP9和HDL-C水平反弹,CTRP5和TG水平降低,IR改善。此外,CLAB可能作为一种有希望的非药物干预,通过调节肠道微生物群和改善脂质和葡萄糖代谢来缓解抑郁症相关的代谢失调。
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引用次数: 0
Thermal Versus Ultrasound Inactivation of Bifidobacterium animalis subsp. lactis BB-12: Functional Implications for Postbiotics. 动物双歧杆菌亚种的热与超声失活。乳酸BB-12:对后生物制剂的功能影响。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-10 DOI: 10.1007/s12602-025-10876-w
Nimet Marsak, Ecem Akan, Adem Yavaş, Zafer Erbay

This study evaluated the viability and functional properties of postbiotics derived from Bifidobacterium animalis subsp. lactis BB-12 following thermal and ultrasound processing. Sixteen processing conditions were tested, including heat treatments (65-95 °C for 5-90 min) and ultrasound amplitudes (98-320 μm for 15-60 min). Postbiotics were assessed for antimicrobial activity, probiotic-stimulating effects on Lacticaseibacillus casei 431 and Bifidobacterium spp. cultures, and their capacity to enhance short-chain fatty acid (SCFA) production. Complete inactivation was achieved under all heat treatment conditions, whereas only 11 ultrasound conditions resulted in total loss of culturability, indicating greater microbial resistance at lower intensities or shorter exposures. Discrepancies between flow cytometry and colony counting in ultrasound-treated samples suggest the presence of viable but non-culturable cells (VBNC), highlighting the limitations of culture-based viability assessments, especially when assessing VBNC states. Postbiotics from heat-treatment showed significantly stronger probiotic-enhancing effects, with growth increases up to 2.9-fold for L. casei and 37.7-fold for Bifidobacterium spp. compared to controls, alongside greater antimicrobial activity, especially against Enterococcus faecalis. Both processing methods significantly increased SCFA levels (p < 0.05). In conclusion, it can be said that heat treatment was more effective than ultrasound in producing biologically active postbiotics from B. lactis BB-12. The enhanced functional properties observed in heat-inactivated preparations underscore thermal processing as a robust method for postbiotic production. These findings highlight the potential of heat-derived postbiotics as active bioingredients for microbiota-targeted functional foods and nutraceuticals. Further in vivo validation and standardization efforts are needed to fully demonstrate their therapeutic potential and support regulatory approval.

本研究评估了动物双歧杆菌亚种衍生的后生物制剂的生存能力和功能特性。经过热和超声处理的乳酸BB-12。测试了16种加工条件,包括热处理(65-95°C, 5-90 min)和超声振幅(98-320 μm, 15-60 min)。研究评估了益生后制剂的抗菌活性、对干酪乳杆菌431和双歧杆菌培养物的益生菌刺激作用,以及它们促进短链脂肪酸(SCFA)产生的能力。在所有热处理条件下都实现了完全失活,而只有11种超声条件导致培养性完全丧失,这表明在较低强度或较短的暴露下,微生物耐药性更强。超声处理的样品中,流式细胞术和菌落计数之间的差异表明存在活细胞但不可培养细胞(VBNC),这突出了基于培养的活力评估的局限性,特别是在评估VBNC状态时。热处理后的益生菌显示出明显更强的益生菌增强作用,与对照组相比,干酪乳杆菌的生长增加了2.9倍,双歧杆菌的生长增加了37.7倍,同时抗菌活性更强,特别是对粪肠球菌。两种处理方法均显著提高了SCFA水平(p
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引用次数: 0
Dietary Saccharomyces Cerevisiae and Pseudomonas Aeruginosa Enhance Growth Performance, Nutrient Utilization, Intestinal Morphology, and Hemato-Biochemical Profile in Red Tilapia (Oreochromis sp.). 饲料中添加酿酒酵母菌和铜绿假单胞菌可提高红罗非鱼的生长性能、营养物质利用、肠道形态和血液生化特征。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-10 DOI: 10.1007/s12602-025-10880-0
Remy Ntakirutimana, K M Mujeeb Rahiman, Jishnu Kunjappan, Megha Lovejan, Aishath Ali Farhath, M P Jijina, A R Nikhila Khana, K Nandhakumar, K V Neethu, Anatole Bigirimana

The search for effective probiotic alternatives to antibiotics in aquaculture has increased in recent years. This study evaluated the dietary supplementation of Saccharomyces cerevisiae and Pseudomonas aeruginosa on growth, feed utilization, intestinal morphology, and health of red tilapia (Oreochromis sp.) over a 60-day feeding trial. Fish were fed diets containing three graded levels of S. cerevisiae (1, 2, and 4 g kg⁻¹ feed) or P. aeruginosa (1, 2, and 3 mL kg⁻¹ feed), and compared with a control group. Both probiotics enhanced performance and physiological responses compared with the control. The highest inclusion levels (4 g kg⁻¹ for S. cerevisiae and 3 mL kg⁻¹ for P. aeruginosa) yielded superior growth rate, feed efficiency, and intestinal villus development, accompanied by improved muscle protein content and stable hematological profiles. No adverse effects were detected in water quality or fish health. These findings highlight the potential of S. cerevisiae and a non-pathogenic P. aeruginosa strain as promising probiotics for red tilapia culture, demonstrating complementary effects on nutrient utilization and physiological condition.

近年来,在水产养殖中寻找抗生素的有效益生菌替代品的工作有所增加。本研究通过60天的饲养试验,评估了饲粮中添加酿酒酵母和铜绿假单胞菌对红罗非鱼生长、饲料利用、肠道形态和健康状况的影响。鱼被喂食含有三个等级的酿酒葡萄球菌(1、2和4克千克毒血症)或铜绿假葡萄球菌(1、2和3毫升千克毒血症)的饲料,并与对照组进行比较。与对照组相比,这两种益生菌都提高了生产性能和生理反应。最高的添加量(酿酒葡萄球菌4克千克(毒血症)和铜绿假单胞菌3毫升千克(毒血症))产生了更高的生长速度、饲料效率和肠绒毛发育,同时改善了肌肉蛋白质含量和稳定的血液学特征。未发现对水质或鱼类健康有不良影响。这些发现凸显了酿酒葡萄球菌和非致病性铜绿假单胞菌作为红罗非鱼养殖益生菌的潜力,表明它们在营养利用和生理条件方面具有互补作用。
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引用次数: 0
Effect of Limosilactobacillus reuteri SBC5-3 Cell-free Supernatant on Growth Performance, Intestinal Inflammation, and Microbiota in Weaned Piglets Challenged With Enterotoxigenic Escherichia coli K88. 罗伊氏乳酸杆菌SBC5-3无细胞上清液对产肠毒素大肠杆菌K88攻毒断奶仔猪生长性能、肠道炎症和微生物群的影响
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-10 DOI: 10.1007/s12602-025-10882-y
Le Xu, Guoying Bao, Shiyu Chen, Chen Liu, Xiannian Zi, Minjie Hu, Qinghua Liu, Qiuye Lin, Zhenhui Cao

The progressive implementation of worldwide restrictions on antibiotic use has necessitated the development of sustainable alternatives for controlling bacterial diarrhea in piglets. This study investigated the effects of a cell-free supernatant (CFS) from Limosilactobacillus reuteri SBC5-3 on growth performance, intestinal inflammation, and microbiota in weaned piglets challenged with enterotoxigenic Escherichia coli K88 (ETEC K88). Forty-eight piglets were randomly assigned to four groups: a control group (CON, basal diet + physiological saline), an ETEC-challenged group (ETEC K88, basal diet + ETEC K88), a prevention group (CFS + ETEC K88, basal diet with CFS supplementation + ETEC K88), and a CFS group (CFS, basal diet with CFS supplementation + physiological saline). The CFS was administered orally at 5 mL/kg body weight for 7 days prior to challenge. Results showed that CFS pretreatment significantly reduced diarrhea rate and fecal scores, improved average daily gain, and alleviated ETEC K88-induced villus damage and lymphocyte infiltration in the jejunum and ileum. Moreover, CFS downregulated the expression of intestinal mucosal pro-inflammatory cytokines (IL1B, IL6, TNFA) and suppressed the activation of the NF-κB pathway by reducing phosphorylation of TAK1, IκBα, and p65. Intestinal microbiota analysis revealed that CFS increased the abundance of Lactobacillus and reduced Escherichia-Shigella. These findings demonstrate that L. reuteri SBC5-3 CFS effectively mitigates ETEC K88-induced diarrhea and intestinal inflammation by inhibiting NF-κB signaling and modulating intestinal microbiota, highlighting its potential as an antibiotic alternative in piglet production.

随着世界范围内对抗生素使用限制的逐步实施,有必要开发可持续的替代品来控制仔猪细菌性腹泻。本试验研究了罗伊氏乳酸杆菌SBC5-3无细胞上清液(CFS)对产肠毒素大肠杆菌K88 (ETEC K88)攻毒断奶仔猪生长性能、肠道炎症和微生物群的影响。48头仔猪随机分为4组:对照组(CON,基础饲粮+生理盐水)、ETEC挑战组(ETEC K88,基础饲粮+ ETEC K88)、预防组(CFS + ETEC K88,基础饲粮中添加CFS + ETEC K88)和CFS组(CFS,基础饲粮中添加CFS +生理盐水)。攻毒前7天,以5 mL/kg体重的剂量口服CFS。结果显示,CFS预处理显著降低腹泻率和粪便评分,提高平均日增重,减轻ETEC k88诱导的空肠和回肠绒毛损伤和淋巴细胞浸润。此外,CFS通过降低TAK1、i -κB α和p65的磷酸化,下调肠黏膜促炎因子(IL1B、IL6、TNFA)的表达,抑制NF-κB通路的激活。肠道菌群分析显示,CFS增加了乳酸菌的丰度,减少了志贺氏杆菌。这些研究结果表明,罗伊氏乳杆菌SBC5-3 CFS通过抑制NF-κB信号传导和调节肠道微生物群,有效减轻ETEC k88诱导的腹泻和肠道炎症,突出了其作为仔猪生产抗生素替代品的潜力。
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引用次数: 0
Characterization and Biofungicide Potential of a Novel Antifungal defensin, K4CBP6, from Solanum lycopersicum L. 一种新型抗真菌防御素K4CBP6的鉴定及生物杀真菌潜力
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-08 DOI: 10.1007/s12602-025-10865-z
Rebeka Papp, Péter Poór, Zalán Czékus, Györgyi Váradi, Zoltán Kele, Attila Borics, Gábor Bende, Kata Horváti, Gábor K Tóth, László Galgóczy, Liliána Tóth

The global rise in resistance to chemical fungicides and their strict regulation by the EU, has created an urgent need for alternative antifungal strategies in agriculture. Plant defensins represent promising alternatives owing to their broad-spectrum antifungal activity, structural stability, and low toxicity to mammalian cells and plants. In this study, we identified and characterized a novel antifungal defensin, K4CBP6, from Solanum lycopersicum L., along with its γ-core peptide derivatives, K4CBP6γ1 and K4CBP6γ2, as potential biofungicide agents. Protein database mining revealed a widespread distribution of K4CBP6 homologs within the Solanaceae family. Recombinant K4CBP6 (rK4CBP6) was successfully produced using a Komagataella phaffii-based expression system, while K4CBP6γ1 and K4CBP6γ2 were chemically synthesized. Structural analyses via electrospray ionization mass spectrometry and electronic circular dichroism spectroscopy confirmed a cysteine-stabilized α-helix β-strand folded structure for rK4CBP6. In vitro susceptibility assays demonstrated that both rK4CBP6 and K4CBP6γ2 exhibited antifungal activity against major tomato pathogens, Botrytis cinerea, Cladosporium herbarum, and Fusarium oxysporum with minimum inhibitory concentrations ranging from 12.5 to 25 µg ml-1. Furthermore, neither rK4CBP6 nor K4CBP6γ2 exhibited cytotoxic effects on mammalian cell lines or adverse effects in animal and plant model systems even at concentrations of 200 and 400 µg ml-1. Proof-of-concept experiments on tomato plants and fruits confirmed their protective efficacy against B. cinerea and C. herbarum. These findings highlight the potential of rK4CBP6 and K4CBP6γ2 as sustainable biofungicide candidates for plant disease management, owing to their antifungal efficacy both in vitro and in planta, along with their lack of cytotoxic effects.

全球对化学杀菌剂的耐药性上升以及欧盟对其的严格监管,迫切需要在农业中寻找替代的抗真菌策略。植物防御素具有广谱抗真菌活性、结构稳定性和对哺乳动物细胞和植物的低毒性,是一种很有前途的替代品。在这项研究中,我们从茄属番茄中鉴定并鉴定了一种新的抗真菌防御素K4CBP6及其γ-核心肽衍生物K4CBP6γ1和K4CBP6γ2,作为潜在的生物杀菌剂。蛋白质数据库挖掘显示,K4CBP6同源物在茄科植物中广泛分布。重组蛋白K4CBP6 (rK4CBP6)通过基于法非氏Komagataella的表达系统成功表达,同时化学合成了K4CBP6γ1和K4CBP6γ2。通过电喷雾电离质谱分析和电子圆二色光谱分析,证实rK4CBP6具有半胱氨酸稳定的α-螺旋- β-链折叠结构。体外药敏试验表明,rK4CBP6和k4cbp6 γ - 2对番茄主要病原菌灰霉病菌(Botrytis cinerea)、草本枝孢菌(Cladosporium herbarum)和尖孢镰刀菌(Fusarium oxysporum)均具有抑制活性,最小抑制浓度为12.5 ~ 25µg ml-1。此外,即使rK4CBP6和K4CBP6γ2浓度分别为200和400µg ml-1,也不会对哺乳动物细胞系产生细胞毒性作用,也不会对动物和植物模型系统产生不良影响。在番茄植株和果实上进行了概念验证实验,证实了其对番茄灰霉病菌和植物芽孢杆菌的保护作用。这些发现突出了rK4CBP6和K4CBP6γ2作为植物病害管理的可持续生物杀菌剂候选物的潜力,因为它们在体外和植物体内都具有抗真菌功效,而且它们没有细胞毒性作用。
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引用次数: 0
Therapeutic Potential of Antimicrobial Peptide Scymicrosin7-26 against the Emerging Pathogen Acinetobacter ursingii Isolated from Litopenaeus vannamei. 抗菌肽Scymicrosin7-26对凡纳滨对虾新发病原体杜氏不动杆菌的治疗潜力
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-05 DOI: 10.1007/s12602-025-10866-y
Ying Wang, Hanxiao Li, Hua Hao, Ying Zhou, Fangyi Chen, Ke-Jian Wang

The Pacific white shrimp (Litopenaeus vannamei) aquaculture faces emerging threats from novel pathogens and escalating antibiotic resistance. This study successfully isolated and identified the pathogenic bacterium Acinetobacter ursingii strain 31C2 from diseased L. vannamei using an integrated approach combining microbiological, biochemical, and molecular techniques. The pathogenicity of this strain was confirmed in L. vannamei and marine medaka (Oryzias melastigma) infection models, exhibiting a strong dose-dependent mortality, with median lethal doses (LD₅₀) of 2.83 × 10⁴ CFU/g shrimp and 2.58 × 10⁶ CFU/fish, respectively. Infection caused severe hepatopancreatic necrosis (tubular deformation and epithelial vacuolation) and intestinal villi destruction. Antimicrobial susceptibility testing revealed that the 31C2 strain was resistant to tetracycline and azithromycin. To identify effective agents targeting this strain, the antimicrobial peptide Scymicrosin7-26 (derived from Scylla paramamosain) was evaluated. The peptide had potent antibacterial activity against A. ursingii 31C2 in vitro (MIC: 3-6 µM). In vivo application significantly enhanced survival of L. vannamei and O. melastigma infected with 31C2 by 30% and 20%, respectively. Treatment drastically reduced bacterial loads in the hepatopancreas and intestine, restored tissue integrity, and modulated the immune response by suppressing the hyperactivation of the Toll and IMD pathways and their downstream transcription factors, dorsal and relish, while upregulating penaeidin3 and propo expression. This study identified A. ursingii as an emerging shrimp pathogen and validated Scymicrosin7-26 as a promising antibiotic-free therapeutic for disease control in aquaculture.

凡纳滨对虾(Litopenaeus vannamei)水产养殖面临着来自新型病原体和不断升级的抗生素耐药性的新威胁。本研究采用微生物学、生化和分子技术相结合的综合方法,成功地从患病的凡纳美乳杆菌中分离和鉴定了致病细菌ursingacinetobacter ursingii菌株31C2。该菌株的致病性在L. vannamei和海洋medaka (Oryzias melastigma)感染模型中得到证实,表现出很强的剂量依赖性死亡率,致死剂量中位数(LD₅0)分别为2.83 × 10⁴CFU/g虾和2.58 × 10⁶CFU/g鱼。感染引起严重的肝胰脏坏死(小管变形和上皮空泡化)和肠绒毛破坏。药敏试验显示31C2菌株对四环素和阿奇霉素均耐药。为了确定针对该菌株的有效药物,我们对抗菌肽Scymicrosin7-26(源自Scylla paramamosain)进行了评估。该肽在体外(MIC: 3 ~ 6µM)对猪链球菌31C2具有较强的抑菌活性。体内应用31C2后,凡纳梅l.s annamei和褪黑o.s melastigma的存活率分别提高30%和20%。治疗显著降低了肝胰脏和肠道中的细菌负荷,恢复了组织完整性,并通过抑制Toll和IMD通路及其下游转录因子dorsal和回味因子的过度激活,同时上调penaeidin3和propo的表达,调节了免疫反应。本研究确定了一种新型的ursingi虾病原体,并验证了Scymicrosin7-26是一种很有前景的水产养殖疾病控制的无抗生素治疗药物。
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引用次数: 0
Genome-Resolved in Silico Analysis of Poultry and Swine Lactobacillales Provides a Data-Driven Framework for Elucidating Metabolic Complementary interactions in Multi-Strain Probiotics. 家禽和猪乳酸杆菌基因组解析的计算机分析为阐明多菌株益生菌代谢互补相互作用提供了数据驱动的框架。
IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-12-05 DOI: 10.1007/s12602-025-10846-2
João Victor Dos Anjos Almeida, Mauro de Medeiros Oliveira, Taís Mayumi Kuniyoshi, Fernando Moisés Mamani Sanca, Carlos Miguel Nóbrega Mendonça, Carlos Emílio Cabrera Matajira, Ana Luiza Louvisi, Ricardo Pinheiro de Souza Oliveira, Alessandro de Mello Varani

Probiotics are live microorganisms that provide health benefits to the host by improving digestion, enhancing nutrient absorption, and modulating the immune system. Among them, lactic acid bacteria are known for producing vitamins and short-chain fatty acids, both essential for intestinal health. In this in silico study, we performed high-fidelity (PacBio HiFi) whole-genome sequencing and comprehensive comparative genomic analysis of five Lactobacillales strains (Enterococcus lactis, Enterococcus mundtii, Ligilactobacillus agilis, Limosilactobacillus reuteri, Limosilactobacillus vaginalis) isolated from the intestinal microbiota of chickens and pigs. The assembled genomes ranged from 1.8 to 2.8 Mb, with more than 98% completeness and less than 1.31% contamination. Taxonomic classification, presence of antimicrobial resistance genes, bacteriocin biosynthetic potential, carbohydrate-active enzyme repertoires and vitamin biosynthesis pathways, and capacity to degrade plant polysaccharides were investigated. Functional characterization identified 65 families of carbohydrate-active enzymes, with E. mundtii presenting the greatest diversity (43 families) and absolute number (100 terms) of enzymes. Metabolic reconstruction suggested functional specialization among strains, with xylooligosaccharide degradation exclusive to E. mundtii and pectin utilization limited to E. lactis. Genes related to the biosynthesis of B-complex vitamins, including riboflavin, folate, and menaquinone, showed heterogeneous and complementary distribution among strains. These findings suggest the potential for metabolic complementarity and cross-feeding, where metabolites produced by one strain serve as precursors for biosynthetic pathways in others. Collectively, these genome-resolved insights offer a data-driven framework for designing multi-strain probiotics aimed at improving intestinal health and feed efficiency in poultry and swine.

益生菌是一种活的微生物,通过改善消化、增强营养吸收和调节免疫系统,为宿主提供健康益处。其中,乳酸菌以生产维生素和短链脂肪酸而闻名,这两种物质对肠道健康都至关重要。在这项计算机研究中,我们对从鸡和猪肠道微生物群中分离的5株乳酸杆菌(乳酸肠球菌、蒙氏肠球菌、柔韧性乳酸乳酸杆菌、罗伊氏乳酸乳酸杆菌、阴道乳酸乳酸杆菌)进行了高保真(PacBio HiFi)全基因组测序和综合比较基因组分析。组装的基因组在1.8 ~ 2.8 Mb之间,完整性大于98%,污染小于1.31%。研究了细菌的分类、耐药基因的存在、细菌素的生物合成潜力、碳水化合物活性酶谱和维生素的生物合成途径以及降解植物多糖的能力。功能鉴定鉴定出65个碳水化合物活性酶家族,其中孟氏酵母(E. mundtii)具有最多的多样性(43个家族)和绝对数量(100个)。代谢重建表明菌株之间的功能特化,低聚木糖的降解仅限于E. mundtii,果胶的利用仅限于E. lactis。与核黄素、叶酸、甲基萘醌等b族复合维生素生物合成相关的基因在菌株间呈现异质性和互补性分布。这些发现表明了代谢互补和交叉喂养的潜力,其中一种菌株产生的代谢物可以作为其他菌株生物合成途径的前体。总的来说,这些基因组解析的见解为设计旨在改善家禽和猪肠道健康和饲料效率的多菌株益生菌提供了数据驱动的框架。
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引用次数: 0
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Probiotics and Antimicrobial Proteins
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