Rapid evolution of hyaluronan synthase to improve hyaluronan production and molecular mass in Bacillus subtilis

Linpei Zhang; Hao Huang; Hao Wang; Jian Chen; Guocheng Du; Zhen Kang

doi:10.1007/s10529-016-2193-1

Rapid evolution of hyaluronan synthase to improve hyaluronan production and molecular mass in Bacillus subtilis

Biotechnol Lett. 2016 Dec;38(12):2103-2108. doi: 10.1007/s10529-016-2193-1. Epub 2016 Aug 26.

Authors

Linpei Zhang¹, Hao Huang¹, Hao Wang¹, Jian Chen^{1

2}, Guocheng Du^{1

3

2}, Zhen Kang^{4

5

6}

Affiliations

¹ The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
² Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, Jiangsu, China.
³ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China.
⁴ The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China. zkang@jiangnan.edu.cn.
⁵ The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China. zkang@jiangnan.edu.cn.
⁶ Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, Jiangsu, China. zkang@jiangnan.edu.cn.

PMID: 27565668
DOI: 10.1007/s10529-016-2193-1

Abstract

Objectives: To improve the production and molecular mass of the glycosaminoglycan hyaluronan (HA) in Bacillus subtilis by engineering hyaluronan synthase (HAS) from Streptococcus zooepidemicus.

Results: By mutating regions within HAS intracellular domains, five positive variants exhibiting higher HA production (from 1.22 to 2.24 g l^-1) and molecular mass values (from 1.20 to 1.36 × 10⁶ Da) were constructed and characterized. Overexpression of the V5 variant and the genes tuaD and glmU increased HA production and molecular mass to 2.8 g l^-1 and 2.4 × 10⁶ Da, respectively.

Conclusions: This study provides a novel strategy for improving HA production and its molecular mass.

Keywords: Bacillus subtilis; Combinatorial evolution; Hyaluronan; Hyaluronan synthase; Molecular mass.

MeSH terms

Bacillus subtilis / enzymology*
Bacillus subtilis / metabolism*
Glucuronosyltransferase / genetics
Glucuronosyltransferase / metabolism
Hyaluronan Synthases
Hyaluronic Acid / biosynthesis*
Molecular Weight

Substances

Hyaluronic Acid
Glucuronosyltransferase
Hyaluronan Synthases