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马蜂肠道菌的分离鉴定及产消化酶功能菌株的筛选
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国家自然科学基金面上项目(42177406);辽宁省自然科学基金面上项目(2023-MS-319)


Isolation and identification of gut bacteria from Vespa mandarinia Smith and screening of digestive enzyme-producing strains
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    摘要:

    【背景】马蜂(Vespa mandarinia Smith)可以防治多种田间害虫,还具有药用价值,其肠道菌群结构和功能还有待研究。【目的】获得马蜂肠道可培养细菌并筛选出具有产消化酶功能的菌株,为理解肠道菌对宿主的影响机理及功能菌株的利用提供科学依据和研究材料。【方法】采用传统细菌分离培养法获得马蜂肠道菌,结合形态学以及16S rRNA基因序列分析进行鉴定;利用水解圈法分别筛选产蛋白酶、脂肪酶、淀粉酶和纤维素酶菌株;通过测量水解圈D与菌落直径d的比值,比较不同细菌的产酶能力。【结果】在马蜂肠道中共分离出6属10种细菌,其中芽孢杆菌属5种,肠球菌属、葡萄球菌属、明串珠菌属、乳球菌属和不动杆菌属各1种。从获得的61个菌株中筛选出6个具有产消化酶功能的菌株。其中,苏云金芽孢杆菌V44具有产蛋白酶、淀粉酶、脂肪酶和纤维素酶4种消化酶的能力;粪肠球菌V6具有产淀粉酶、蛋白酶和脂肪酶3种消化酶的能力;蜡样芽孢杆菌V43具有产蛋白酶、淀粉酶和纤维素酶3种消化酶的能力;粪肠球菌V20、蜡样芽孢杆菌V19和维德曼氏芽孢杆菌V22均具有产蛋白酶的能力。【结论】马蜂肠道细菌资源较丰富,部分有产消化酶的功能,可帮助马蜂消化食物,对宿主健康有一定影响。本研究筛选的6个菌株都能产蛋白酶,其中菌株V43和V44分别具有最强产淀粉酶和脂肪酶的能力,是可进一步开发利用的肠道功能菌株资源。

    Abstract:

    [Background] Vespa mandarinia Smith can control a variety of field pests and has medicinal values. The structure and function of gut bacteria in V. mandarinia remain to be studied. [Objective] To obtain the culturable bacteria from the gut of V. mandarinia and screen out the bacterial strains capable of producing digestive enzyme, so as to provide a basis and research materials for understanding the influence of gut bacteria on the host and for utilization of the functional bacterial strains. [Methods] The gut bacteria of V. mandarinia were isolated by the culture method, and 16S rRNA gene sequencing was employed to identify the strains. Halo-forming method was adopted to screen out the strains producing protease, lipase, amylase, and cellulase, and the ratio of halo diameter (D) to bacterial colony diameter (d) was used to compare the strain capability of producing digestive enzymes. [Results] A total of 10 species of bacteria belonging to 6 genera were isolated from the gut of V. mandarinia, including 5 species of Bacillus, 1 species of Enterococcus, Staphylococcus, Leuconostoc, Lactococcus, and Acinetobacter, respectively. Six strains producing digestive enzymes were screened out from the 61 strains obtained. Among them, B. thuringiensis V44 had the ability to produce protease, amylase, lipase, and cellulase. E. faecalis V6 was capable of producing amylase, protease, and lipase. B. cereus V43 can produce protease, amylase, and cellulase. E. faecalis V20, B. cereus V19, and B. wiedmannii V22 demonstrated the ability to produce protease. [Conclusion] The gut bacteria resources of V. mandarinia were abundant, and some strains were capable of producing digestive enzymes, which can facilitate the digestion and affection of health of V. mandarinia. All the 6 strains screened out in this study could produce protease. Strains V43 and V44 showed the strongest ability to produce amylase and lipase, respectively, which could be further developed as gut functional strain resources.

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何雨薇,陈芳敏,夏琬婷,刘娴,黄晨晨. 马蜂肠道菌的分离鉴定及产消化酶功能菌株的筛选[J]. 微生物学通报, 2023, 50(6): 2624-2634

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  • 收稿日期:2022-09-09
  • 录用日期:2022-12-27
  • 在线发布日期: 2023-06-05
  • 出版日期: 2023-06-25
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