食品科学 ›› 2024, Vol. 45 ›› Issue (8): 87-95.doi: 10.7506/spkx1002-6630-20230714-161

• 生物工程 • 上一篇    下一篇

产酯酶格氏乳球菌的筛选、鉴定与基因组注释

陈聪,邹伟,汤秀娟,陈晓松,吴成泽   

  1. (1.四川轻化工大学生物工程学院,四川 宜宾 644005;2.酿酒生物技术及应用四川省重点实验室,四川 宜宾 644005;3.乐山职业技术学院,四川 乐山 614000)
  • 出版日期:2024-04-25 发布日期:2024-05-01
  • 基金资助:
    泸州老窖研究生创新基金项目(LJCX2022-2);四川轻化工大学校级大学生创新创业训练计划项目(cx2022126); 四川轻化工大学研究生创新基金项目(Y2023234)

Screening, Identification and Genome Annotation of Esterase-Producing Lactococcus garvieae

CHEN Cong, ZOU Wei, TANG Xiujuan, CHEN Xiaosong, WU Chengze   

  1. (1. College of Bioengineering, Sichuan University of Science & Engineering, Yibin 644005, China; 2. Liquor Brewing Biotechnology and Application Key Laboratoray of Sichuan Province, Yibin 644005, China; 3. Leshan Vocational and Technical College, Leshan 614000, China)
  • Online:2024-04-25 Published:2024-05-01

摘要: 本研究从浓香型白酒大曲进行初筛和复筛,得到一株高酯酶活力的菌株S5-4,对其全基因组测序、酯酶注释和代谢通路分析。该菌株的最大透明圈D/d比值(细菌透明圈直径与菌落直径之比)达到了2.27±0.02,经鉴定为格氏乳球菌(Lactococcus garvieae)。对其形态及生理生化特性进行鉴定,并对其产酯酶的酶活力进行测定,其产酯酶的活力为15.74 U/mL,在35 ℃厌氧发酵15 d能够合成乙酸乙酯(0.345 0±0.160 0)g/L、乳酸乙酯(0.298 3±0.230 0)g/L。S5-4基因组测序与注释结果显示,其基因组大小为2 191 113 bp,GC相对含量为37.99%,预测得到了1 486 个基因,其中4 个核糖体RNA、69 个转移RNA和34 个非编码RNA基因。S5-4基因组共注释出11 种产酯酶基因,其中包括磷酸酯酶(胞内酶5 个和细胞膜酶2 个)、酰基辅酶A脱酰酶(胞内酶)、丙酸三酯酶(胞外酶)、羧酸酯酶(胞外酶)和木聚糖酯酶(胞内酶)的基因。基于功能基因组注释构建了S5-4以淀粉和半乳糖为碳源生产乙酸乙酯和乳酸乙酯代谢路径,与实验结果相符。根据菌株产酯酶特性,可用作在白酒酿造中酯化液的生物催化剂,或者将其作为大曲增香的强化剂。

关键词: 格氏乳球菌;基因组信息;代谢路径重建;酯酶

Abstract: In this study, a strain, named S5-4, with high esterase activity was obtained from Nongxiangxing Daqu by primary and secondary screening, and subjected to whole genome sequencing, esterase annotation and metabolic pathway analysis. The strain exhibited a maximal ratio of transparent zone diameter to colony diameter (D/d) of 2.27 ± 0.02, and it was identified as Lactococcus garvieae. Its morphological, physiological, and biochemical characteristics were determined, and its esterase activity was measured to be 15.74 U/mL. This strain produced (0.345 0 ± 0.160 0) g/L of ethyl acetate and (0.298 3 ± 0.230 0) g/L of ethyl lactate after anaerobic fermentation at 35 ℃ for 15 days. The genome size of S5-4 was determined to be 2 191 113 bp in length, with a GC content of 37.99%. A total of 1 486 genes, including 4 rRNA genes, 69 tRNA genes, and 34 ncRNA genes were predicted. In total, 11 esterase genes were annotated in the genome of S5-4 including phosphatases (five intracellular ones and two membrane ones), acyl-CoA dehydrogenase (intracellular), triacylglycerol lipase (extracellular), carboxylase (extracellular), and hemicellulose esterase (intracellular). Based on functional genomic annotation, the metabolic pathways to produce ethyl acetate and ethyl lactate from starch and galactose as carbon sources were constructed and found to be consistent with the experimental results. Given its ability to produce esterase, the strain could be used as a biocatalyst for esterification in baijiu fermentation or as a flavor enhancer in Daqu production.

Key words: Lactococcus garvieae; genomic information; metabolic pathway reconstruction; esterase

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