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The Function of β-1,4-Glucuronosyltransferase WelK in the Sphingan WL Gum Biosynthesis Process in Marine Sphingomonas sp. WG

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Abstract

The marine-derived polysaccharide WL gum produced by Sphingomonas sp. WG showed commercial utility potential in ink, food, and oil industries. A β-1,4-glucuronosyltransferase WelK was predicted to catalyze the transfer of glucuronic acid from UDP-glucuronic acid to glucosyl-α-pyrophosphorylpolyprenol intermediate in the WL gum biosynthesis process. Its function was evaluated by bioinformatical analysis, gene knocking out, and overexpressing strategies. Compared to the wild strain, the WL gum production and broth viscosity of the mutant ∆welK were decreased by 71.5% and 99.2% when cultured for 48 h. The gene disruption led to the failure of product preparation. Homologous expression of welK in the native organism can effectively improve WL gum production. When glucose concentration was 6.7%, the WL gum production by the welK-overexpressing strain cultured for 60 h and 84 h reached 32.65 and 43.13 g/L, 134.1%, and 114% of the wild strain. The polysaccharide composition and qRT-PCR analysis showed that the glucuronic acid content was closely related to the expression level of welK. Thus, WelK was proved to play a critical role in the WL gum synthesis and will be an attractive target for metabolic engineering. Our experiment provided a genetic manipulation method for the functional characterization of genes in Sphingomonas sp. WG.

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Funding

This work was financially supported by the National Natural Science Foundation of China (31800075 and U1805234), 863 Program (2015AA020925), Natural Science Foundation of Fujian Province of China (2019J01264), Minjiang Scholar (2013A13), startup fund for high-level talent at Fujian Normal University (004828), Fundamental Research Funds for the Central Universities (18CX02124A), and the Development Fund of State Key Laboratory of Heavy Oil Processing (20CX02202A).

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Authors and Affiliations

  1. State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering and Biotechnology, China University of Petroleum (East China), 66 Changjiang West Road, Qingdao, 266580, People’s Republic of China

    Hui Li, Kehui Li, Zhongrui Guo, Han Xue, Jing Li, Sixue Ji, Jiqian Wang & Hu Zhu

  2. Engineering Research Center of Industrial Biocatalysis, Fujian Province Higher Education Institutes, College of Chemistry and Materials Science, Fujian Normal University, 32 Shangsan Road, Fuzhou, 350007, People’s Republic of China

    Hu Zhu

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  1. Hui Li
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  2. Kehui Li
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  3. Zhongrui Guo
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Correspondence to Jiqian Wang or Hu Zhu.

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Li, H., Li, K., Guo, Z. et al. The Function of β-1,4-Glucuronosyltransferase WelK in the Sphingan WL Gum Biosynthesis Process in Marine Sphingomonas sp. WG. Mar Biotechnol 23, 39–50 (2021). https://doi.org/10.1007/s10126-020-09998-9

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