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UGT708S6 from Dendrobium catenatum, catalyzes the formation of flavonoid C-glycosides. 来自铁皮石斛的 UGT708S6 催化黄酮 C-糖苷的形成。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-19 DOI: 10.1186/s12896-024-00923-9
Luyao Yu, Kun He, Yu Wu, Kai Hao, Yun Wang, Jinbo Yao, Yuxue Zhao, Qiaoxian Yu, Yanghui Shen, Mengxuan Chen, Ke Xu, Xinfeng Zhang, Lei Zhang

Background: Dendrobium catenatum is a perennial herb of the genus Dendrobium orchidaceae. It has been known as "Golden Grass, Soft Gold" since ancient times with effects of strengthening the body, benefiting the stomach, generating body fluid, nourishing Yin and clearing internal heat. The flowers of D. catenatum have anti-oxidation, immune regulation and other biological activities. The composition analysis of flowers showed that flavonoid glycosides were significantly accumulated in floral tissue. However, in the flowers of D. catenatum, there was only one case of the UDP-glycosyltransferase (UGT) responsible for the glycosylation of flavonoids has been reported.

Result: In this study, a new UGT (named UGT708S6) was cloned from D. catenatum flowers rich in O-glycosides and C-glycosides, and its function and biochemical properties were characterized. Through homology comparison and molecular docking, we identified the key amino acid residues affecting the catalytic function of UGT708S6. The glycosyltransferase UGT708S6 was characterized and demonstrated C-glycosyltransferase (CGT) activity in vitro assay using phloretin and 2-hydroxynaringenin as sugar acceptors. The catalytic promiscuity assay revealed that UGT708S6 has a clear sugar donor preference, and displayed O-glycosyltransferase (OGT) activity towards luteolin, naringenin and liquiritigenin. Furthermore, the catalytic characteristics of UGT708S6 were explored, shedding light on the structural basis of substrate promiscuity and the catalytic mechanism involved in the formation of flavonoid C-glycosides. R271 was a key amino acid residue site that sustained the catalytic reaction. The smaller binding pocket resulted in the production of new O-glycosides and the reduction of C-glycosides. This highlighted the importance of the binding pocket in determining whether C-glycosides or O-glycosides were produced.

Conclusions: The findings suggest that UGT708S6 holds promise as a new glycosyltransferase for synthesizing flavonoid glycosides and offer valuable insights for further understanding the catalytic mechanisms of flavonoid glycosyltransferases.

背景介绍铁皮石斛为兰科石斛属多年生草本植物。自古就有 "黄金草、软黄金 "的美誉,具有强身健体、益胃生津、滋阴清热的功效。泽泻花具有抗氧化、免疫调节等生物活性。花的成分分析表明,黄酮苷类在花组织中明显积累。然而,在 D. catenatum 的花中,负责类黄酮糖基化的 UDP-糖基转移酶(UGT)仅有一例报道:结果:本研究从富含 O-糖苷和 C-糖苷的 D. catenatum 花中克隆了一种新的 UGT(命名为 UGT708S6),并对其功能和生化特性进行了表征。通过同源性比较和分子对接,我们确定了影响 UGT708S6 催化功能的关键氨基酸残基。我们对糖基转移酶 UGT708S6 进行了表征,并在体外实验中以毛果芸香素和 2-hydroxynaringenin 为糖受体证明了其 C-糖基转移酶(CGT)活性。催化杂合性测定显示,UGT708S6 对糖受体有明显的偏好,对木犀草素、柚皮素和琉璃苣甙元具有 O 型糖基转移酶(OGT)活性。此外,研究人员还探讨了 UGT708S6 的催化特性,揭示了底物杂合性的结构基础以及形成黄酮类 C-糖苷的催化机理。R271 是维持催化反应的关键氨基酸残基位点。较小的结合口袋导致产生新的 O 型糖苷和减少 C 型糖苷。这凸显了结合袋在决定产生 C-糖苷还是 O-糖苷方面的重要性:研究结果表明,UGT708S6有望成为合成黄酮苷的新型糖基转移酶,并为进一步了解黄酮糖基转移酶的催化机理提供了有价值的见解。
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引用次数: 0
Evaluating the impact of Cold plasma on Seedling Growth properties, seed germination, and soybean antioxidant enzyme activity. 评估冷等离子体对幼苗生长特性、种子发芽和大豆抗氧化酶活性的影响。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-14 DOI: 10.1186/s12896-024-00921-x
Khadijeh Sayahi, Amir Hossein Sari, Aidin Hamidi, Bahareh Nowruzi, Farshid Hassani

Cold atmospheric pressure plasma (CAP) has garnered significant attention in recent years for its potential applications in biomedical, environmental, and agricultural fields. Cold plasma treatment exhibits a variety of effects in agricultural applications, including impacts on seed germination and seedling growth; however, further research is required. Soybean serves as a fundamental source of nutrients for both animals and humans. Soybean seeds possess impermeable and thick testae, which results in prolonged germination times and suboptimal germination rates. The soybeans exhibit low uniformity. As a result, poor crop establishment and yield reduction are inevitable outcomes. Therefore, the purpose of this study was to examine the effects of Iranian soybean cultivars, such as Sari, Saba, Arian, Katoul, and Williams, on seedling growth properties, seed germination, and antioxidant enzyme activity, using argon at time intervals of 30, 60, 180, 300, and 420 s. Cold plasma treatment significantly enhanced germination potential from 1.18 to 66.97%, germination index from 0.50 to 60.09%, germination rate from 1.78 to 32.17%, seedling length from 2.70 cm to 78.13 cm, root length from 2.87 cm to 56.13 cm, and seedling dry weight from 1.80 g to 36.63 g. Additionally, CAT activity increased from 0.88- to 4.40-fold, SOD activity from 0.86- to 5.89-fold, and APX activities from 0.40- to 4.01-fold compared to the control treatment. The findings indicated that the samples exhibited optimal results at treatment durations of 60 and 180 s. The influence of plasma on the antioxidant responses of seedlings, seed germination, and growth characteristics was contingent upon the duration of treatment. Cold plasma, when applied for an appropriate duration, may enhance soybean seedling growth characteristics and seed germination.

近年来,冷等离子体(CAP)因其在生物医学、环境和农业领域的潜在应用而备受关注。冷等离子体处理在农业应用中表现出多种效果,包括对种子发芽和幼苗生长的影响;然而,还需要进一步的研究。大豆是动物和人类的基本营养来源。大豆种子具有不透气的厚种皮,导致发芽时间延长,发芽率不理想。大豆的均匀度很低。因此,作物生长不良和减产是不可避免的结果。因此,本研究的目的是研究伊朗大豆栽培品种,如 Sari、Saba、Arian、Katoul 和 Williams,在 30、60、180、300 和 420 秒的时间间隔内使用氩气对幼苗生长特性、种子萌发和抗氧化酶活性的影响。此外,与对照处理相比,CAT 活性提高了 0.88 至 4.40 倍,SOD 活性提高了 0.86 至 5.89 倍,APX 活性提高了 0.40 至 4.01 倍。研究结果表明,样品在处理持续时间为 60 秒和 180 秒时效果最佳。在适当的时间内使用冷等离子体,可以提高大豆幼苗的生长特性和种子发芽率。
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引用次数: 0
Isolation and identification of antifungal, antibacterial and nematocide agents from marine bacillus gottheilii MSB1. 从海洋杆菌 MSB1 中分离和鉴定抗真菌、抗细菌和杀线虫剂。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-13 DOI: 10.1186/s12896-024-00920-y
Ahmed S Shehata, Marwa A Samy, Sherien E Sobhy, Aida M Farag, Ibrahim M El-Sherbiny, Ahmed A Saleh, Elsayed E Hafez, Mamdouh Abdel-Mogib, Haiam M Aboul-Ela

Pathogenic fungi employ numerous strategies to colonize plants, infect them, reduce crop yield and quality, and cause significant losses in agricultural production. The increasing use of chemical pesticides has led to various ecological and environmental issues, including the emergence of resistant weeds, soil compaction, and water pollution, all negatively impacting agricultural sustainability. Additionally, the extensive development of synthetic fungicides has adverse effects on animal and human health, prompting the exploration of alternative approaches and green strategies for phytopathogen control. Microorganisms living in sponges represent a promising source of novel bioactive secondary metabolites, potentially useful in developing new nematicidal and antimicrobial agents. This study focuses on extracting bioactive compounds from endosymbiotic bacteria associated with the marine sponge Hyrtios erect sp. (collected from NIOF Station, Hurghada, Red Sea, Egypt) using various organic solvents. Bacillus sp. was isolated and identified through 16 S rRNA gene sequencing. The biocidal activity of Bacillus gotheilii MSB1 extracts was screened against plant pathogenic bacteria, fungi, and nematodes. The n-butanol extract showed significant potential as a biological fungicide against Alternaria alternata and Fusarium oxysporum. Both n-hexane and ethyl acetate extracts exhibited negative impacts against the plant pathogenic bacteria Erwinia carotovora and Ralstonia solanacearum, whereas the n-butanol extract had a positive effect. Regarding nematicidal activity, ethyl acetate and n-butanol extracts demonstrated in-vitro activity against the root-knot nematode Meloidogyne incognita, which causes serious vegetable crop diseases, but the n-hexane extract showed no positive effects. The findings suggest that bioactive compounds from endosymbiotic bacteria associated with marine sponges, particularly B. gotheilii MSB1, hold significant potential as alternative biological control agents against plant pathogens. The n-butanol extract, in particular, displayed promising biocidal activities against various plant pathogenic fungi, bacteria, and nematodes. These results support further exploration and development of such bioactive compounds as sustainable, environmentally friendly alternatives to synthetic pesticides and fungicides in agricultural practices.

病原真菌采用多种策略定殖植物,感染植物,降低作物产量和质量,给农业生产造成重大损失。化学杀虫剂使用量的不断增加导致了各种生态和环境问题,包括抗性杂草的出现、土壤板结和水污染,所有这些都对农业的可持续发展产生了负面影响。此外,合成杀菌剂的广泛开发也对动物和人类健康产生了不利影响,这促使人们探索植物病原体控制的替代方法和绿色策略。生活在海绵中的微生物是新型生物活性次生代谢物的重要来源,可能有助于开发新型杀线虫剂和抗菌剂。本研究的重点是利用各种有机溶剂从与海洋海绵直立藻(Hyrtios erect sp.,采集自埃及红海赫尔格达的 NIOF 站)相关的内共生细菌中提取生物活性化合物。通过 16 S rRNA 基因测序,分离并鉴定了芽孢杆菌。筛选了枯草芽孢杆菌 MSB1 提取物对植物病原菌、真菌和线虫的杀菌活性。正丁醇提取物作为生物杀真菌剂对交替交替孢霉和氧孢镰刀菌具有显著的潜力。正己烷和乙酸乙酯提取物对植物病原菌 Erwinia carotovora 和 Ralstonia solanacearum 都有负面影响,而正丁醇提取物则有正面影响。在杀线虫活性方面,乙酸乙酯和正丁醇提取物对导致严重蔬菜作物病害的根结线虫(Meloidogyne incognita)表现出体外活性,但正己烷提取物没有表现出积极作用。研究结果表明,与海洋海绵相关的内共生细菌(尤其是 B. gotheilii MSB1)的生物活性化合物具有作为替代生物防治剂防治植物病原体的巨大潜力。特别是正丁醇提取物,对各种植物病原真菌、细菌和线虫都有很好的生物杀灭活性。这些结果支持进一步探索和开发此类生物活性化合物,作为农业实践中合成杀虫剂和杀真菌剂的可持续、环保型替代品。
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引用次数: 0
Analysis of hsa_circ_0136256 as a biomarker for fibrosis in systemic sclerosis. 分析作为系统性硬化症纤维化生物标志物的 hsa_circ_0136256。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-13 DOI: 10.1186/s12896-024-00910-0
Xiaolin Sun, Baoyue Wang, Lili Ding, Yongfu Wang, Mingguo Xu

Background: Exploration of whether circRNAs in the skin of systemic sclerosis (SSc) model mice interact with 4E-BP1 protein to mediate the mTOR signaling pathway to regulate SSc fibrosis is crucial to identify homologous human circRNAs as markers to guide the diagnosis and treatment of SSc.

Methods: C57BL/6 mice aged 6-8 weeks and weighing approximately 20 g were subcutaneously injected with bleomycin (BLM) to establish an SSc model. High-throughput sequencing was used to screen the differentially expressed circRNA in the skin of SSc model mice and control mice. RNA immunoprecipitation and RNA pulldown confirmed the interaction between circRNA and 4E-BP1 protein. SSc model mice were treated with empty plasmid (OE-NC), overexpression plasmid of mmu_circ_0005372 (OE-circ_0005372), interference plasmid of mmu_circ_0005372 (sh-circ5372), mutant plasmid of mmu_circ_0005372 (circ5372-MT), mTOR activator (MHY1485), mTOR inhibitor (omipalisib), or JAK1/2 inhibitor (ruxolitinib). Sections of mouse skin tissue were stained with Hematoxylin and eosin and Masson's stain. The collagen volume fraction (CVF) was calculated as CVF = area of blue collagen/total area with ImageJ. The correlation between homologous human circRNAs and clinical data was analyzed.

Results: Compared to the control group, 21,839 circRNAs were upregulated and 27, 946 circRNAs were downregulated in the skin tissue of mice in the SSc model group. Among them was mmu_circ_0005372, which is derived from the FZD3 gene, is closely related to fibrosis, and is involved in the mTOR signaling pathway. Hsa_circ_0136256 was identified as the homologous human circRNA of mmu_circ_0005372. RT-qPCR confirmed that the expression of mmu_circ_0005372 was significantly reduced in the skin tissue of SSc mice, and the expression of hsa_circ_0136256 was significantly reduced in the peripheral blood mononuclear cells of patients with SSc. The interaction between mmu_circ_0005372 and 4E-BP1 protein was inhibited in the skin tissue of SSc model mice. The results showed that the CVF of OE-circ_0005372 group was significantly lower than that of the sh-circ5372, circ5372-MT, and MHY1485 groups, indicating that OE-circ5372 significantly improved skin fibrosis in the SSc mice. ROC curve analysis was performed on hsa_circ_0136256 (AUC = 0.719, P = 0.035). The expression of hsa_circ_0136256 was negatively correlated with COL IV, RDW-SD, and RDW-CV, and positively correlated with VC, PLT, and PCT. The results suggested that hsa_circ_0136256 may have important roles in the clinical diagnosis of SSc.

Conclusion: Mmu_circ_0005372 and homologous human hsa_circ_0136256 may be biomarkers and therapeutic targets for SSc fibrosis.

背景:探索系统性硬化症(SSc)模型小鼠皮肤中的circRNA是否与4E-BP1蛋白相互作用,介导mTOR信号通路以调控SSc纤维化,这对于确定同源的人类circRNA作为指导SSc诊断和治疗的标志物至关重要:方法:对年龄为6-8周、体重约20克的C57BL/6小鼠皮下注射博莱霉素(BLM),建立SSc模型。采用高通量测序筛选SSc模型小鼠和对照小鼠皮肤中差异表达的circRNA。RNA免疫沉淀和RNA pulldown证实了circRNA与4E-BP1蛋白之间的相互作用。用空质粒(OE-NC)、mmu_circ_0005372的过表达质粒(OE-circ_0005372)、mmu_circ_0005372的干扰质粒(sh-circ5372)、mmu_circ_0005372的突变质粒(circ5372-MT)、mTOR激活剂(MHY1485)、mTOR抑制剂(奥米帕利西)或JAK1/2抑制剂(鲁索利替尼)处理SSc模型小鼠。小鼠皮肤组织切片用苏木精、伊红和马森氏染色法染色。胶原体积分数(CVF)用ImageJ计算,即CVF=蓝色胶原面积/总面积。分析了同源人类 circRNA 与临床数据之间的相关性:结果:与对照组相比,SSc模型组小鼠皮肤组织中有21839个circRNA上调,27946个circRNA下调。其中 mmu_circ_0005372 源自 FZD3 基因,与纤维化密切相关,参与 mTOR 信号通路。Hsa_circ_0136256 被鉴定为 mmu_circ_0005372 的同源人类 circRNA。RT-qPCR 证实,在 SSc 小鼠的皮肤组织中,mmu_circ_0005372 的表达明显降低,而在 SSc 患者的外周血单核细胞中,hsa_circ_0136256 的表达也明显降低。在 SSc 模型小鼠的皮肤组织中,mmu_circ_0005372 与 4E-BP1 蛋白的相互作用受到抑制。结果显示,OE-circ_0005372 组的 CVF 明显低于 sh-circ5372、circ5372-MT 和 MHY1485 组,表明 OE-circ5372 能显著改善 SSc 小鼠的皮肤纤维化。对 hsa_circ_0136256 进行了 ROC 曲线分析(AUC = 0.719,P = 0.035)。hsa_circ_0136256 的表达与 COL IV、RDW-SD 和 RDW-CV 呈负相关,与 VC、PLT 和 PCT 呈正相关。结果表明,hsa_circ_0136256在SSc的临床诊断中可能具有重要作用:结论:Mmu_circ_0005372 和同源人类 hsa_circ_0136256 可能是 SSc 纤维化的生物标志物和治疗靶点。
{"title":"Analysis of hsa_circ_0136256 as a biomarker for fibrosis in systemic sclerosis.","authors":"Xiaolin Sun, Baoyue Wang, Lili Ding, Yongfu Wang, Mingguo Xu","doi":"10.1186/s12896-024-00910-0","DOIUrl":"10.1186/s12896-024-00910-0","url":null,"abstract":"<p><strong>Background: </strong>Exploration of whether circRNAs in the skin of systemic sclerosis (SSc) model mice interact with 4E-BP1 protein to mediate the mTOR signaling pathway to regulate SSc fibrosis is crucial to identify homologous human circRNAs as markers to guide the diagnosis and treatment of SSc.</p><p><strong>Methods: </strong>C57BL/6 mice aged 6-8 weeks and weighing approximately 20 g were subcutaneously injected with bleomycin (BLM) to establish an SSc model. High-throughput sequencing was used to screen the differentially expressed circRNA in the skin of SSc model mice and control mice. RNA immunoprecipitation and RNA pulldown confirmed the interaction between circRNA and 4E-BP1 protein. SSc model mice were treated with empty plasmid (OE-NC), overexpression plasmid of mmu_circ_0005372 (OE-circ_0005372), interference plasmid of mmu_circ_0005372 (sh-circ5372), mutant plasmid of mmu_circ_0005372 (circ5372-MT), mTOR activator (MHY1485), mTOR inhibitor (omipalisib), or JAK1/2 inhibitor (ruxolitinib). Sections of mouse skin tissue were stained with Hematoxylin and eosin and Masson's stain. The collagen volume fraction (CVF) was calculated as CVF = area of blue collagen/total area with ImageJ. The correlation between homologous human circRNAs and clinical data was analyzed.</p><p><strong>Results: </strong>Compared to the control group, 21,839 circRNAs were upregulated and 27, 946 circRNAs were downregulated in the skin tissue of mice in the SSc model group. Among them was mmu_circ_0005372, which is derived from the FZD3 gene, is closely related to fibrosis, and is involved in the mTOR signaling pathway. Hsa_circ_0136256 was identified as the homologous human circRNA of mmu_circ_0005372. RT-qPCR confirmed that the expression of mmu_circ_0005372 was significantly reduced in the skin tissue of SSc mice, and the expression of hsa_circ_0136256 was significantly reduced in the peripheral blood mononuclear cells of patients with SSc. The interaction between mmu_circ_0005372 and 4E-BP1 protein was inhibited in the skin tissue of SSc model mice. The results showed that the CVF of OE-circ_0005372 group was significantly lower than that of the sh-circ5372, circ5372-MT, and MHY1485 groups, indicating that OE-circ5372 significantly improved skin fibrosis in the SSc mice. ROC curve analysis was performed on hsa_circ_0136256 (AUC = 0.719, P = 0.035). The expression of hsa_circ_0136256 was negatively correlated with COL IV, RDW-SD, and RDW-CV, and positively correlated with VC, PLT, and PCT. The results suggested that hsa_circ_0136256 may have important roles in the clinical diagnosis of SSc.</p><p><strong>Conclusion: </strong>Mmu_circ_0005372 and homologous human hsa_circ_0136256 may be biomarkers and therapeutic targets for SSc fibrosis.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"91"},"PeriodicalIF":3.5,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11562351/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142614254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Enhanced vitamin B12 production by isolated Bacillus strains with the application of response surface methodology. 应用响应面方法提高分离芽孢杆菌菌株的维生素 B12 产量。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-12 DOI: 10.1186/s12896-024-00919-5
Rania M M Abdel-Baki, Marwa N Ahmed, Olfat S Barakat, Galal M Khalafalla

Background: Vitamin B12 is a crucial B-group vitamin, first isolated from the liver due to its role in combating pernicious anemia. It is distinguished by its unique and complex structure, which makes its chemical synthesis challenging and expensive. Consequently, vitamin B12 is alternatively obtained through microbial fermentations. Molasses, an affordable and safe agro-industrial waste, can be used as a carbon source for vitamin B12 production, offering a cost-effective alternative to expensive sugars in the production medium.

Results: A total of 87 yeast, actinomycete, and bacterial isolates were screened for vitamin B12 production, with 15 isolates showing high productivity. Bacillus isolates were selected for further analysis using MALDI-TOF and molecular identification. These isolates were identified as four strains of Bacillus subtilis (MZ08, JT10, BY11, and JT17), one strains of Bacillus sp. (CB09), and one strain of Peribacillus acanthi (MZ01). Genetic circuits associated with vitamin B12 production were demonstrated in a closely related strain of Peribacillus acanthi MZ01 strain. Three strains (MZ01, MZ08, and JT17) were selected for further evaluation of vitamin B12 productivity under different sugar types (glucose, sucrose, fructose, lactose, and galactose) and varying inoculum sizes. The inoculum size significantly impacted vitamin B12 production, with an increase from 5 to 10% enhancing yields. The ability of the strains to produce vitamin B12 varied depending on the type of sugar used. Peribacillus acanthi MZ01 strain showed the highest productivity and subsequently, selected for optimizing vitamin B12 production conditions using response surface methodology. Furthermore, the optimized conditions were then applied to molasses-based medium to achieve high vitamin B12 yields by MZ01 strain.

Conclusion: In this study, Peribacillus acanthi was characterized for the first time as a vitamin B12 producer, demonstrating high productivity among various tested strains. The optimization of production conditions using response surface methodology, further enhanced vitamin B12 yields, showcasing the strain's efficiency in microbial fermentations. This research also highlights the potential of using molasses as a cost-effective alternative carbon source, significantly reducing production costs.

背景:维生素 B12 是一种重要的 B 族维生素,最初是从肝脏中分离出来的,因为它具有防治恶性贫血的作用。其独特而复杂的结构使其化学合成具有挑战性且成本高昂。因此,维生素 B12 可通过微生物发酵获得。糖蜜是一种既经济又安全的农用工业废料,可用作生产维生素 B12 的碳源,为生产培养基中昂贵的糖类提供了一种具有成本效益的替代品:结果:共筛选出 87 个酵母、放线菌和细菌分离物用于生产维生素 B12,其中 15 个分离物显示出较高的生产率。筛选出的芽孢杆菌分离物通过 MALDI-TOF 和分子鉴定进行了进一步分析。这些分离菌株被鉴定为四株枯草芽孢杆菌(MZ08、JT10、BY11 和 JT17)、一株芽孢杆菌(CB09)和一株尖头弧菌(MZ01)。与维生素 B12 生产相关的基因回路在一株密切相关的尖头弧菌(Peribacillus acanthi)MZ01 菌株中得到了证实。为了进一步评估不同糖类(葡萄糖、蔗糖、果糖、乳糖和半乳糖)和不同接种量下维生素 B12 的产量,选取了三个菌株(MZ01、MZ08 和 JT17)。接种物的大小对维生素 B12 的产量有很大影响,接种物从 5%增加到 10%会提高产量。菌株生产维生素 B12 的能力因糖的种类而异。Acanthi Peribacillus MZ01 菌株表现出最高的生产率,因此被选中利用响应面方法优化维生素 B12 的生产条件。此外,将优化后的条件应用于以糖蜜为基础的培养基,MZ01 菌株的维生素 B12 产量很高:本研究首次鉴定了尖头嗜酸乳杆菌(Peribacillus acanthi)作为维生素 B12 生产者的特性,在各种测试菌株中显示出较高的生产率。利用响应面方法优化生产条件,进一步提高了维生素 B12 的产量,展示了该菌株在微生物发酵中的高效性。这项研究还凸显了使用糖蜜作为具有成本效益的替代碳源的潜力,从而大大降低了生产成本。
{"title":"Enhanced vitamin B<sub>12</sub> production by isolated Bacillus strains with the application of response surface methodology.","authors":"Rania M M Abdel-Baki, Marwa N Ahmed, Olfat S Barakat, Galal M Khalafalla","doi":"10.1186/s12896-024-00919-5","DOIUrl":"10.1186/s12896-024-00919-5","url":null,"abstract":"<p><strong>Background: </strong>Vitamin B<sub>12</sub> is a crucial B-group vitamin, first isolated from the liver due to its role in combating pernicious anemia. It is distinguished by its unique and complex structure, which makes its chemical synthesis challenging and expensive. Consequently, vitamin B<sub>12</sub> is alternatively obtained through microbial fermentations. Molasses, an affordable and safe agro-industrial waste, can be used as a carbon source for vitamin B<sub>12</sub> production, offering a cost-effective alternative to expensive sugars in the production medium.</p><p><strong>Results: </strong>A total of 87 yeast, actinomycete, and bacterial isolates were screened for vitamin B<sub>12</sub> production, with 15 isolates showing high productivity. Bacillus isolates were selected for further analysis using MALDI-TOF and molecular identification. These isolates were identified as four strains of Bacillus subtilis (MZ08, JT10, BY11, and JT17), one strains of Bacillus sp. (CB09), and one strain of Peribacillus acanthi (MZ01). Genetic circuits associated with vitamin B<sub>12</sub> production were demonstrated in a closely related strain of Peribacillus acanthi MZ01 strain. Three strains (MZ01, MZ08, and JT17) were selected for further evaluation of vitamin B<sub>12</sub> productivity under different sugar types (glucose, sucrose, fructose, lactose, and galactose) and varying inoculum sizes. The inoculum size significantly impacted vitamin B<sub>12</sub> production, with an increase from 5 to 10% enhancing yields. The ability of the strains to produce vitamin B<sub>12</sub> varied depending on the type of sugar used. Peribacillus acanthi MZ01 strain showed the highest productivity and subsequently, selected for optimizing vitamin B<sub>12</sub> production conditions using response surface methodology. Furthermore, the optimized conditions were then applied to molasses-based medium to achieve high vitamin B<sub>12</sub> yields by MZ01 strain.</p><p><strong>Conclusion: </strong>In this study, Peribacillus acanthi was characterized for the first time as a vitamin B<sub>12</sub> producer, demonstrating high productivity among various tested strains. The optimization of production conditions using response surface methodology, further enhanced vitamin B<sub>12</sub> yields, showcasing the strain's efficiency in microbial fermentations. This research also highlights the potential of using molasses as a cost-effective alternative carbon source, significantly reducing production costs.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"90"},"PeriodicalIF":3.5,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11555979/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142614270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Antioxidant, neuroprotective, and neuroblastoma cells (SH-SY5Y) differentiation effects of melanins and arginine-modified melanins from Daedaleopsis tricolor and Fomes fomentarius. 三色堇和福美双的黑色素及精氨酸修饰黑色素的抗氧化、神经保护和神经母细胞瘤细胞(SH-SY5Y)分化作用。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-11 DOI: 10.1186/s12896-024-00918-6
Hoang Anh Thu Nguyen, Thien Phu Ho, Debby Mangelings, Ann Van Eeckhaut, Yvan Vander Heyden, Hanh T M Tran

Background: Microbial melanins possess a broad spectrum of biological activities. However, there is little understanding of their neuroprotective and neuronal cell differentiation properties. This study aimed to extract, purify, and modify melanins from two medicinal fungi (Daedaleopsis tricolor and Fomes fomentarius), and to evaluate their antioxidant activity, as well as their cell protective ability against neurotoxins. In addition, the study also investigated the feasibility of combining melanins or modified melanins with retinoic acid (RA) to induce neuronal differentiation.

Methods: Melanin was extracted and purified using alkaline acid-based methods. Antioxidant activities and neuroprotective effects were evaluated using the DPPH (1,1-diphenyl-2-picrylhydrazyl) and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assays, respectively. In addition, morphological changes of SH-SY5Y cells were recorded by using a Pannoramic MIDI scanner.

Results: All melanins and arginine-modified melanins displayed mild DPPH scavenging activities, which were statistically lower than that of ascorbic acid (p < 0.05). In terms of neuroprotection, both melanins and arginine-modified melanins exhibited significant cell protection against H2O2 after 24 h exposure (p < 0.05). Notably, there is no significant difference between F. fomentarius melanin and its modified form as they both increased cell viability by about 20%. Contrarily, while D. tricolor melanin enhanced the cell viability with 16%, its modified form increased the cell viability with 21%. These activities, however, are significantly lower than the positive control (N-acetylcysteine, p < 0.05). Regarding MPTP, only the arginine-modified melanins of the two fungi significantly protected the cells after 24 h exposure to the toxin (p < 0.05). Specifically, F. fomentarius and D. tricolor modified melanins enhanced the cell viability with 10.2% and 11.1%, respectively, whereas that of the positive control was 13.2%. Interestingly, combining RA (10 µM) with 20 µg/mL of either F. fomentarius, or especially D. tricolor arginine-modified melanin, significantly promoted neuroblastoma cell differentiation into mature neuronal cells compared to using RA alone (p < 0.05).

Conclusions: The arginine-modified melanins of D. tricolor and F. fomentarius have potential for neuroprotection against Parkinsonian neurotoxins. In addition, the arginine-modified melanin of D. tricolor may serve as an excellent material for research in neuroblastoma treatment.

背景:微生物黑色素具有广泛的生物活性。然而,人们对它们的神经保护和神经细胞分化特性了解甚少。本研究旨在从两种药用真菌(Daedaleopsis tricolor 和 Fomes fomentarius)中提取、纯化和修饰黑色素,并评估它们的抗氧化活性以及对神经毒素的细胞保护能力。此外,该研究还探讨了将黑色素或经修饰的黑色素与维甲酸(RA)结合以诱导神经元分化的可行性:方法:使用基于碱性酸的方法提取和纯化黑色素。方法:采用碱性酸法提取纯化黑色素,分别用 DPPH(1,1-二苯基-2-苦基肼)和 MTT(3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑)法评估抗氧化活性和神经保护作用。此外,还使用 Pannoramic MIDI 扫描仪记录了 SH-SY5Y 细胞的形态变化:结果:所有黑色素和精氨酸修饰黑色素都显示出轻微的 DPPH 清除活性,在统计学上低于抗坏血酸(p < 0.05)。在神经保护方面,黑色素和精氨酸修饰的黑色素在暴露于 H2O2 24 小时后都表现出明显的细胞保护作用(p < 0.05)。值得注意的是,F. fomentarius 黑色素与其修饰形式之间没有明显差异,它们都能使细胞存活率提高约 20%。相反,三色堇黑色素提高了 16% 的细胞活力,而其改良形式提高了 21% 的细胞活力。不过,这些活性明显低于阳性对照(N-乙酰半胱氨酸,p < 0.05)。至于 MPTP,只有两种真菌的精氨酸修饰黑色素能在细胞暴露于毒素 24 小时后明显保护细胞(p < 0.05)。具体来说,F. fomentarius 和 D. tricolor 改良黑色素分别提高了细胞存活率 10.2% 和 11.1%,而阳性对照的提高率为 13.2%。有趣的是,与单独使用 RA 相比,将 RA(10 µM)与 20 µg/mL 的 F. fomentarius 或 D. tricolor 精氨酸修饰黑色素结合使用,能显著促进神经母细胞瘤细胞分化为成熟的神经元细胞(p < 0.05):结论:D. tricolor 和 F. fomentarius 的精氨酸修饰黑色素具有针对帕金森神经毒素的神经保护潜力。此外,三色堇的精氨酸修饰黑色素可作为神经母细胞瘤治疗研究的绝佳材料。
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引用次数: 0
Fertility protective effects of Brillantaisia patula leaf extract against cyclophosphamide-induced ovarian damage in Wistar rats. 斑鸠菊叶提取物对环磷酰胺诱导的 Wistar 大鼠卵巢损伤的生育保护作用
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-08 DOI: 10.1186/s12896-024-00916-8
Olalekan Bukunmi Ogunro, Bankole Emmanuel Ofeniforo

Background: The primary indication of infertility is the incapacity to conceive, and in females, the majority of instances of female infertility stem from ovulation disorders. This study evaluated the female fertility-enhancing effects and safety of aqueous leaf extract of Brillantaisia patula (ALEBP) in a cyclophosphamide (CYP) model of sterility in Wistar rats.

Method: Sixty-six female rats randomly allotted to six groups (n = 11) were administered with the appropriate regimen for 21 days and then mated with male rats. Group 1 (control) received distilled water. Groups 2-6 were treated with a single dose (200 mgkg- 1 body weight) of cyclophosphamide intraperitoneally and, in addition, received the same volume (0.5 mL) of distilled water, 18, 36, 72 mgkg- 1 body weight of ALEBP and 200 mg per body weight of vitamin C orally. Mating lasted 11 days; on day 20, the female Wistar rats were sacrificed. Data were analysed using One-way Analysis of Variance (ANOVA) followed by Dunett's posthoc analysis, and GraphPad (at p < 0.05).

Results: Results herein showed that ALEBP significantly (p < 0.05) increased the diminution in activities/levels of glutathione peroxidase (GPx), reduced glutathione (GSH), total antioxidant capacity (TAC), cholesterol, alkaline phosphatase (ALP), acid phosphatase (ACP), estrogen (ES), and luteinising hormone (LH) induced by cyclophosphamide. ALEBP further reversed the increased level of malondialdehyde (MDA), tumour necrosis factor-α (TNFα), interleukin 8 (IL-8), and follicle-stimulating hormone (FSH) caused by cyclophosphamide (p < 0.05). In addition, ALEBP, while it significantly increased the cyclophosphamide-induced reduction in the number of implantations in each animal, the total number of viable fetuses, the total number of corpora lutea, and the fertility index, also significantly reduced the number of fetal resorptions in each animal and pre-implantation loss that was increased by cyclophosphamide. Moreover, the cyclophosphamide-induced degenerative and necrotic changes in the ovarian cells and uterus were reversed by ALEBP.

Conclusions: Considered as a whole, the aqueous leaf extract of Brillantaisia patula reversed oxidative stress and inflammatory side effects of cyclophosphamide, preserving ovarian function and fertility in the rats. This may suggest its exploration as a safe agent against toxic side effects of chemotherapy and fertility-related disorders of the uterus and ovary.

背景:不孕症的主要表现是无法受孕,而女性不孕症大多源于排卵障碍。本研究评估了在环磷酰胺(CYP)不育大鼠模型中,板蓝根水叶提取物(ALEBP)对雌性生育力的增强作用和安全性:将66只雌性大鼠随机分为6组(n = 11),按适当的方案给药21天,然后与雄性大鼠交配。第 1 组(对照组)饮用蒸馏水。第 2-6 组腹腔注射单剂量(200 毫克/千克-1 体重)环磷酰胺,此外还口服相同体积(0.5 毫升)的蒸馏水、18、36、72 毫克/千克-1 体重的 ALEBP 和 200 毫克/体重的维生素 C。交配持续 11 天;第 20 天,雌性 Wistar 大鼠被处死。数据分析采用单因素方差分析(ANOVA)和Dunett事后分析,以及GraphPad(P结果):结果表明,ALEBP 显著(p 结论:从整体上看,ALEBP 对雌激素的影响较小:从整体上看,斑蝥水叶提取物可逆转环磷酰胺的氧化应激和炎症副作用,保护大鼠的卵巢功能和生育能力。这可能表明,它可以作为一种安全的制剂,防止化疗的毒副作用以及与生育有关的子宫和卵巢疾病。
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引用次数: 0
Fabrication of apigenin and adenosine-loaded nanoparticles against doxorubicin-induced myocardial infarction by reducing inflammation and oxidative stress. 制备芹菜素和腺苷负载纳米粒子,通过减少炎症和氧化应激来预防多柔比星诱发的心肌梗死
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-05 DOI: 10.1186/s12896-024-00912-y
Ruixuan Li, Aixia Xu, Ye Chen, Yihui Li, Ru Fu, Weihong Jiang, Xiaogang Li

The study's goals are to fabricate PLGA nanoparticles (PNPs) loaded with apigenin (AP) and adenosine (AD) using a microfluidic preparation method to a standard emulsification method and investigate the possible heart-protective effects of AP-AD PNPs made using the emulsification method. Compared to microfluidics, the emulsification method fabricated small-size nanoparticles, which are better at encapsulating drugs, retaining more drugs, and having a low viscosity for the myocardial infarction (MI) injection. TheMI model was developed using SD rats injected under the skin with 85 mg/kg doxorubicin (DOX) for 2 days. The metabolic results showed that our AP-AD PNPs accelerated the blood flow in rats with MI, which increased the amounts of AP and AD in the circulatory system. This led to significant improvements in the cardiac index and lower amounts of AST, LDH, and CK in the blood. A histopathological study using Hematoxylin&eosin, and TUNEL staining showed that cardiac function had improved and apoptosis had decreased. Moreover, tests that checked the amounts of IL-6, TNF-α, NO, GSH, MDA, and SOD showed that AP-AD PNPs may help treat MI by reducing oxidative stress and inflammation, making it a potentially useful therapeutic approach.

该研究的目的是用微流控制备法和标准乳化法制备负载有芹菜素(AP)和腺苷(AD)的聚乳酸丙烯酸酯(PLGA)纳米粒子(PNPs),并研究用乳化法制备的AP-AD PNPs可能具有的心脏保护作用。与微流控制备法相比,乳化法制备的纳米粒子尺寸更小,更能包裹药物、保留更多药物,而且粘度低,更适合心肌梗死(MI)注射。心肌梗死模型是利用 SD 大鼠皮下注射 85 毫克/千克多柔比星(DOX)2 天后建立的。代谢结果表明,我们的 AP-AD PNPs 加快了心肌梗死大鼠的血流速度,增加了循环系统中的 AP 和 AD 含量。这导致心脏指数明显改善,血液中的 AST、LDH 和 CK 含量降低。使用苏木精和 TUNEL 染色法进行的组织病理学研究表明,心脏功能得到改善,细胞凋亡减少。此外,对 IL-6、TNF-α、NO、GSH、MDA 和 SOD 含量的检测表明,AP-AD PNPs 可通过减少氧化应激和炎症来帮助治疗心肌梗死,是一种潜在的有效治疗方法。
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引用次数: 0
Limonene encapsulated alginate/collagen as antibiofilm drug against Acinetobacter baumannii. 将柠檬烯包裹的海藻酸盐/胶原蛋白作为抗鲍曼不动杆菌的生物膜药物。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-01 DOI: 10.1186/s12896-024-00888-9
Fatemeh-Sadat GholamhosseinTabar Valookolaei, Hossein Sazegar, Leila Rouhi

This work examined the antibacterial and antibiofilm properties of alginate/collagen nanoparticles containing limonene. The multi-drug resistant (MDR) strains were screened, and the morphological features of the produced nanoparticles were determined utilizing SEM, DLS, and FTIR. Additionally, the encapsulation effectiveness, stability, and drug release were assessed. The levels of OmpA and Bap biofilm genes were assessed using qRT-PCR. At the same time, the antibacterial and cytotoxic activities of the nanoparticles were evaluated using well diffusion and MTT techniques, respectively. LAC nanoparticles measuring 300 ± 9.6 nm in size, 83.64 ± 0.19% encapsulation efficiency, and 60-day stability at 4 °C were synthesized. The biological investigation demonstrated that LAC nanoparticles had potent antibacterial capabilities. This was shown by their ability to significantly decrease the transcription of OmpA and Bap biofilm genes at a statistically significant level of p ≤ 0.05. The nanoparticles exhibited reduced antibiotic resistance compared to free limonene and alginate/collagen. Compared to limonene, LAC nanoparticles exhibited negligible cytotoxicity against HEK-293 at doses ranging from 1.56 to 100 µg/mL (p ≤ 0.01). The findings underscore the potential of LAC nanoparticles as a breakthrough in the fight against highly resistant pathogens. The potent antibacterial effects of LAC nanoparticles versus Acinetobacter baumannii (A. baumannii) MDR strains, considered highly resistant pathogens of significant concern, could inspire new strategies in antibacterial research.

本研究考察了含有柠檬烯的海藻酸/胶原蛋白纳米粒子的抗菌和抗生物膜特性。筛选了耐多药(MDR)菌株,并利用扫描电镜、DLS 和傅立叶变换红外光谱测定了所制纳米粒子的形态特征。此外,还对封装效果、稳定性和药物释放进行了评估。利用 qRT-PCR 技术评估了 OmpA 和 Bap 生物膜基因的水平。同时,还分别使用井扩散和 MTT 技术评估了纳米颗粒的抗菌和细胞毒性活性。合成的 LAC 纳米粒子大小为 300 ± 9.6 nm,封装效率为 83.64 ± 0.19%,在 4 °C 下可稳定 60 天。生物学研究表明,LAC 纳米粒子具有强大的抗菌能力。这表现在它们能够显著降低 OmpA 和 Bap 生物膜基因的转录,且 p ≤ 0.05 具有统计学意义。与游离的柠檬烯和海藻酸/胶原相比,纳米粒子表现出更低的抗生素耐药性。与柠檬烯相比,在剂量为 1.56 至 100 µg/mL 时,LAC 纳米粒子对 HEK-293 的细胞毒性可忽略不计(p ≤ 0.01)。这些发现凸显了 LAC 纳米粒子在抗击高抗药性病原体方面的突破潜力。LAC 纳米粒子对鲍曼不动杆菌(A. baumannii)MDR 菌株的强效抗菌作用可激发抗菌研究的新策略。
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引用次数: 0
Fusarium verticillioides pigment: production, response surface optimization, gamma irradiation and encapsulation studies. 轮状镰刀菌色素:生产、响应面优化、伽马射线照射和封装研究。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-10-30 DOI: 10.1186/s12896-024-00909-7
Mai Ali Mwaheb, Yasmeen A Hasanien, Amira G Zaki, Alaa S Abdel-Razek, Laila R Abd Al Halim

Background: Natural pigments are becoming more significant because of the rising cost of raw materials, pollution, and the complexity of synthetic pigments. Compared to synthetic pigments, natural pigments exhibit antimicrobial properties and is less allergic. Pigments from microbial sources could easily be obtained in an inexpensive culture media, produced in high yields, and microbes are capable of producing different colored pigments. Searching for new sources for natural pigments to replace synthetic ones in food applications has become an urgent necessity, but the instability of these compounds is sometimes considered one of the obstacles that reduce their application. Encapsulation provides an ideal solution for natural dye protection through a controlled release strategy. Thus, this study aims at isolation of several soil fungi and subsequent screening their pigment production ability. The chosen pigment-producing fungal strain underwent full identification. The produced pigment was extracted with ethyl acetate and estimated spectrophotometrically. As there is a necessity to obtain a high pigment yield for efficient industrial application, the best production medium was tested, optimum conditions for maximum dye production were also investigated through the response surface methodology, and gamma irradiation was also employed to enhance the fungal productivity. Encapsulation of the produced pigment into chitosan microsphere was tested. The pigment release under different pH conditions was also investigated.

Results: A new strain, Fusarium verticillioides AUMC 15934 was chosen and identified for a violet pigment production process. Out of four different media studied, the tested strain grew well on potato dextrose broth medium. Optimum conditions are initial medium pH 8, 25 °C-incubation temperature, and for 15-day incubation period under shaking state. Moreover, a 400 Gy irradiation dose enhanced the pigment production. Chitosan microsphere loaded by the pigment was successfully prepared and characterized by infrared spectroscopy and scanning electron microscopy.

Conclusion: This irradiated Fusarium strain provides a more economically favorable source for production of a natural violet dye with an optimum productivity, enhanced yield, and improved properties (such as, enhanced stability, controlled release, and bioaccessibility) by encapsulation with chitosan for efficient application in food industry.

背景:由于原材料成本上升、污染和合成颜料的复杂性,天然颜料正变得越来越重要。与合成颜料相比,天然颜料具有抗菌特性,过敏性较低。微生物来源的颜料可以很容易地在廉价的培养基中获得,产量高,而且微生物能够生产不同颜色的颜料。寻找天然色素的新来源以取代食品中的合成色素已成为当务之急,但这些化合物的不稳定性有时被认为是减少其应用的障碍之一。封装技术通过控制释放策略为天然染料的保护提供了理想的解决方案。因此,本研究旨在分离几种土壤真菌,并随后筛选它们的色素生产能力。所选的色素生产真菌菌株经过了全面鉴定。产生的色素用乙酸乙酯提取,并用分光光度法进行估算。由于需要获得较高的色素产量以实现有效的工业应用,因此对最佳生产介质进行了测试,还通过响应面方法研究了染料产量最大化的最佳条件,并采用伽马射线照射来提高真菌的生产率。还测试了将生产的颜料封装到壳聚糖微球中的情况。此外,还研究了不同 pH 值条件下的色素释放情况:结果:选择并鉴定了一株新菌株 Fusarium verticillioides AUMC 15934,用于紫色颜料的生产过程。在所研究的四种不同培养基中,被测菌株在马铃薯葡萄糖肉汤培养基上生长良好。最佳条件是初始培养基 pH 值为 8,培养温度为 25 °C,在振荡状态下培养 15 天。此外,400 Gy 的辐照剂量可提高色素产量。成功制备了负载色素的壳聚糖微球,并通过红外光谱和扫描电子显微镜对其进行了表征:该辐照镰刀菌菌株为生产天然紫色染料提供了一种更经济的来源,通过壳聚糖封装,该染料具有最佳的生产率、更高的产量和更好的性能(如更高的稳定性、可控释放性和生物可及性),可有效地应用于食品工业。
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引用次数: 0
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BMC Biotechnology
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