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Screening, cloning, expression and characterization of new alkaline trehalose synthase from Pseudomonas monteilii and its application for trehalose production.
Journal of Microbiology and Biotechnology ( IF 2.8 ) Pub Date : 2021-08-20 , DOI: 10.4014/jmb.2106.06032
Srisakul Trakarnpaiboon 1 , Benjarat Bunterngsook 1 , Rungtiva Wansuksriand 2 , Verawat Champreda 1
Affiliation  

Trehalose is a non-reducing disaccharide with an increasing demand for applications in food, nutraceutical, and pharmaceutical industries. Single step trehalose production by trehalose synthase (TreS) using maltose as a starting material is a promising alternative process for industrial application due to its simplicity and cost advantage. Pseudomonas monteilii TBRC 1196 was identified using the developed screening method as a potent strain for TreS production. The TreS gene from P. monteilii TBRC 1196 was firstly cloned and expressed in Escherichia coli. Purified recombinant trehalose synthase (PmTreS) had a molecular weight of 76 kDa and showed optimal pH and temperature at 9.0 and 40°C, respectively. The enzyme exhibited >90% residual activity under mesophilic condition under a broad pH range 7-10 for 6 hours. Maximum trehalose yield by PmTreS was 68.1% with low yield of glucose (4%) as a byproduct under optimal conditions, equivalent to productivity of 4.5 g/l/h using enzyme loading of 2 mg/g substrate and high concentration maltose solution (100 g/l) in a lab-scale bioreactor. The enzyme represents a potent biocatalyst for energy-saving trehalose production with potential on inhibiting microbial contamination by alkaline condition.

中文翻译:

蒙特氏假单胞菌新型碱性海藻糖合酶的筛选、克隆、表达、表征及其在海藻糖生产中的应用。

海藻糖是一种非还原性二糖,在食品、保健品和制药行业的应用需求不断增加。以麦芽糖为起始原料,通过海藻糖合酶 (TreS) 单步生产海藻糖,由于其简单性和成本优势,是一种很有前途的工业应用替代工艺。使用开发的筛选方法将假单胞菌 monteilii TBRC 1196 鉴定为 TreS 生产的有效菌株。首次从P. monteilii TBRC 1196 克隆并在大肠杆菌中表达 TreS 基因. 纯化的重组海藻糖合酶 (PmTreS) 的分子量为 76 kDa,最佳 pH 和温度分别为 9.0 和 40°C。该酶在 7-10 的宽 pH 范围和 6 小时的嗜温条件下表现出 >90% 的残留活性。在最佳条件下,PmTreS 的最大海藻糖产量为 68.1%,作为副产物的葡萄糖产量较低 (4%),相当于使用 2 mg/g 底物和高浓度麦芽糖溶液 (100 g / l)在实验室规模的生物反应器中。该酶代表了一种有效的生物催化剂,用于节能海藻糖生产,具有抑制碱性条件引起的微生物污染的潜力。
更新日期:2021-08-20
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