PROTEIN EXPRESSION AND PURIFICATION
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2022
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IF 2.025
Xiaowen Wang,Sibo Chen,Lin Fan,Hao Liu,Si Chen,Xianfeng Ye,Zhoukun Li,Zhongli Cui,Yan Huang
A new glycoside hydrolase family 2 (GH2) β-galactosidase encoding gene galM was cloned from Microvirga sp. strain MC18 and overexpressed in Escherichia coli . The recombinant β-galactosidase GalM showed optimal activity at pH 7.0 and 50 °C, with a stability at pH 6.0–9.0 and 20–40 °C, which are conditions suitable for the diary environment. The K m and V max values for o-nitrophenyl-β- d -galactopyranoside (oNPG) were 1.30 mmol/L and 15.974 μmol/(min·mg), respectively. GalM showed low product inhibition by galactose with a K i of 73.18 mM and high tolerance for glucose that 86.5% activity retained in the presence of 500 mM glucose. It was also stable and active in 20% of methanol, ethanol and isopropanol. In addition, the enzyme activity of GalM was activated significantly over 0–2 mol/L NaCl (1.6–4.3 fold). These favorable properties make GalM a potential candidate for the industrial application.
研究领域:
生物化学
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酶学
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生物技术
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2024
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IF 4.8
Xinyu Yao,Jinying Gao,Lanqiao Wang,Xiaoning Hou,Litao Ge,Xinxin Qin,Jiazhang Qiu,Xuming Deng,Wei Li,Jianfeng Wang
Ethnopharmacology relevance Cananga oil (CO) is derived from the flowers of the traditional medicinal plant, the ylang-ylang tree. As a traditional antidepressant, CO is commonly utilized in the treatment of various mental disorders including depression, anxiety, and autism. It is also recognized as an efficient antibacterial insecticide, and has been traditionally utilized to combat malaria and acute inflammatory responses resulting from bacterial infections both in vitro and in vivo . Aim of the study The objective of this study is to comprehensively investigate the anti- Salmonella activity and mechanism of CO both in vitro and in vivo , with the expectation of providing feasible strategies for exploring new antimicrobial strategies and developing novel drugs. Methods The in vitro antibacterial activity of CO was comprehensively analyzed by measuring MIC, MBC, growth curve, time-killing curve, surface motility, biofilm, and Live/dead bacterial staining. The analysis of the chemistry and active ingredients of CO was conducted using GC-MS. To examine the influence of CO on the membrane homeostasis of Salmonella , we conducted utilizing diverse techniques, including ANS, PI, NPN, ONPG, BCECF-AM, DiSC3(5), and scanning electron microscopy (SEM) analysis. In addition, the antibacterial mechanism of CO was analyzed and validated through metabolomics analysis. Finally, a mouse infection model of Salmonella typhimurium was established to evaluate the toxic side effects and therapeutic effects of CO. Results The antibacterial effect of CO is the result of the combined action of the main chemical components within its six (palmitic acid, α-linolenic acid, stearic acid, benzyl benzoate, benzyl acetate, and myristic acid). Furthermore, CO disrupts the balance of purine metabolism and the tricarboxylic acid cycle (TCA cycle) in Salmonella , interfering with redox processes. This leads to energy metabolic disorders and oxidative stress damage within the bacteria, resulting in bacterial shock, enhanced membrane damage, and ultimately bacterial death. It is worth emphasizing that CO exerts an effective protective influence on Salmonella infection in vivo within a non-toxic concentration range. Conclusion The outcomes indicate that CO displays remarkable anti- Salmonella activity both in vitro and in vivo . It triggers bacterial death by disrupting the balance of purine metabolism and the TCA cycle, interfering with the redox process, making it a promising anti- Salmonella medication.
研究领域:
药理学
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微生物学
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代谢组学
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传染病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）