携带截短型透明质酸合酶的新型枯草芽孢杆菌生产透明质酸及其特性研究
Hyaluronic acid production and characterization by novel Bacillus subtilis harboring truncated Hyaluronan Synthase.
作者信息
Amjad Zanjani Fatemeh Sadat, Afrasiabi Shadi, Norouzian Dariush, Ahmadian Gholamreza, Hosseinzadeh Sara Ali, Fayazi Barjin Alireza, Cohan Reza Ahangari, Keramati Malihe
机构信息
Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, Iran.
Department of Industrial and Environmental Biotechnology, National Institute for Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
出版信息
AMB Express. 2022 Jul 12;12(1):88. doi: 10.1186/s13568-022-01429-3.
Hyaluronic Acid (HA) is a natural biopolymer that has important physiological and industrial applications due to its viscoelastic and hydrophilic characteristics. The responsible enzyme for HA production is Hyaluronan synthase (HAS). Although in vitro structure-function of intact HAS enzyme has been partly identified, there is no data on in vivo function of truncated HAS forms. In the current study, novel recombinant Bacillus subtilis strains harboring full length (RBSFA) and truncated forms of SeHAS (RBSTr4 and RBSTr3) were developed and HA production was studied in terms of titer, production rate and molecular weight (Mw). The maximum HA titer for RBSFA, RBSTr4 and RBSTr3 was 602 ± 16.6, 503 ± 19.4 and 728 ± 22.9 mg/L, respectively. Also, the HA production rate was 20.02, 15.90 and 24.42 mg/L.h, respectively. The findings revealed that RBSTr3 produced 121% and 137% more HA rather than RBSFA and RBSTr4, respectively. More interestingly, the HA Mw was about 60 kDa for all strains which is much smaller than those obtained in prior studies.
透明质酸（HA）是一种天然生物聚合物，因其粘弹性和亲水特性而具有重要的生理和工业应用。负责HA产生的酶是透明质酸合酶（HAS）。尽管完整HAS酶的体外结构-功能已部分确定，但关于截短形式的HAS在体内功能的数据尚无报道。在本研究中，构建了携带全长SeHAS（RBSFA）和截短形式（RBSTr4和RBSTr3）的新型重组枯草芽孢杆菌菌株，并从滴度、产率和分子量（Mw）方面研究了HA的产生。RBSFA、RBSTr4和RBSTr3的最大HA滴度分别为602±16.6、503±19.4和728±22.9mg/L。此外，HA产率分别为20.02、15.90和24.42mg/（L·h）。研究结果表明，RBSTr3产生的HA分别比RBSFA和RBSTr4多121%和137%。更有趣的是，所有菌株的HA Mw约为60kDa, 这比先前研究中获得的Mw小得多。
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