Screening, Cloning, Expression and Characterization of New Alkaline Trehalose Synthase from Pseudomonas monteilii and Its Application for Trehalose Production

• Published in: Journal of microbiology and biotechnology Vol. 31; no. 10; pp. 1455 - 1464
• Main Authors: Trakarnpaiboon, Srisakul, Bunterngsook, Benjarat, Wansuksriand, Rungtiva, Champreda, Verawat
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society for Microbiology and Biotechnology 28.10.2021; 한국미생물·생명공학회
• Subjects: Amino Acid Sequence; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cloning, Molecular; Enzyme Stability; Glucose - metabolism; Glucosyltransferases - genetics; Glucosyltransferases - metabolism; Maltose - metabolism; Pseudomonas - enzymology; Recombinant Proteins - metabolism; Research article; Trehalose - biosynthesis; 생물학; maltose; trehalose synthase; Pseudomonas monteilii; Trehalose
• DOI: 10.4014/jmb.2106.06032
• ISSN: 1017-7825

Trehalose is a non-reducing disaccharide in 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. TBRC 1196 was identified using the developed screening method as a potent strain for TreS production. The TreS gene from TBRC 1196 was first cloned and expressed in . 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 of 7-10 for 6 h. 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 for inhibiting microbial contamination by alkaline condition.