周培华，何 娟，钟文涛，宋 阳*，刘 兰，郑真真，刘 宇

（湖南省产商品质量检验研究院，食品安全监测与预警湖南省重点实验室，长沙410007）

摘要：该文从自然界中选育具有海藻糖生产潜力的菌株，先经具有高筛选压力的初筛培养基获得一定量的菌株，再通过高效液相色谱（High-performance liquid chromatography，HPLC）准确测定目标菌株的海藻糖生成转化效果。通过建立筛选方法，从温泉样本HS-202006211109中筛到一株能产海藻糖的优质菌株HS-202006211109-117，其在45 ℃、6 h下转化麦芽糖生成海藻糖量为3.73 mg/mL，优于文献报道3 h转化生成0.56 g/L，经革兰氏染色、生化鉴定及荧光定量PCR鉴定为耐热芽孢杆菌，该菌株具有较优异的遗传稳定性和环境耐受性等性能。该文选育能高效转化麦芽糖生成海藻糖的高产菌株，为实现海藻糖的生产核心技术自主化提供技术支撑。

关键词：海藻糖；高产菌株；高通量筛选；高效液相色谱

中图分类号：TS201.3 文献标识码：A 文章编号：1674-506X（2024）06-0105-0005

Selective Breeding and Transformation Determination of Trehalose-producing Strain

ZHOU Peihua，HE Juan，ZHONG Wentao，SONG Yang*，LIU Lan，ZHENG Zhenzhen，LIU Yu

（Hunan Provincial Key Laboratory of Food Safety Monitoring and Early Warning，Hunan Provincial Institute of Product and Goods Quality Inspection，Changsha 410007，China）

Abstract：In this paper, the strains with trehalose production potential were selected from the nature, and a certain amount of strains were first obtained by the primary screening medium with high screening pressure, and then the transformation effect of the trehalose production of the target strain was determined by high-performance liquid chromatography (HPLC). A high-quality strain HS-202006211109-117 which can produce trehalose was screened from the hot spring sample HS-202006211109 by the established screening method, and the amount of trehalose produced by it was 3.73 mg/mL at 45 ℃ for 6 h, which was better than that report in the literature as 0.56 g/L by 3 h transformation. After Gram stained, biochemical identification and fluorescence quantitative PCR, the strains were identified as Bacillus thermotolerans. The strains had excellent genetic stability and environmental tolerance. This paper selectively bred high-yield strains that can transform maltose to produce trehalose, provided technical support for the autonomy of key technology for trehalose production.

Keywords：trehalose；high-yield strain；high-throughput screening；high-performance liquid chromatography

doi：10.3969/j.issn.1674-506X.2024.06-015