Temperature-controlled molecular weight of hyaluronic acid produced by engineered Bacillus subtilis
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ORIGINAL RESEARCH PAPER
Temperature-controlled molecular weight of hyaluronic acid produced by engineered Bacillus subtilis Yingying Li . Zhuangzhuang Shi . Yuzhe Shao . Mengmeng Wu . Guoqiang Li . Ting Ma
Received: 15 February 2020 / Accepted: 3 September 2020 Springer Nature B.V. 2020
Abstract To produce high-, medium- and lowmolecular-weight hyaluronic acid (HA) at different temperatures using engineered Bacillus subtilis expressing hyaluronidase (HAase) from leech. By overexpressing the HAase gene hya in the HAproducing strain WmB using temperature-sensitive plasmid pKSV7, the engineered strain WmB-PYh produced HA with different molecular weights (8.61 kDa at 32 C, 0.615 MDa at 42 C, and 6.19 MDa at 47 C). In this study, the molecular weight of HA was regulated by using leech HAase expressed
from a temperature-sensitive plasmid. We thus obtained different molecular weight HAs by using a single bacterial strain at different culture temperatures. Keywords Bacillus subtilis Hyaluronic acid Hyaluronidase Molecular weight Microbial engineering Temperature-sensitive expression
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-03001-0) contains supplementary material, which is available to authorized users. Y. Li Z. Shi Y. Shao M. Wu G. Li T. Ma (&) Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071, China e-mail: [email protected] Y. Li e-mail: [email protected] Z. Shi e-mail: [email protected] Y. Shao e-mail: [email protected] M. Wu e-mail: [email protected] G. Li e-mail: [email protected]
Hyaluronic acid (HA) is a non-sulphated glycosaminoglycan that has been applied in cosmetic, food and pharmaceutical industries (Kogan et al. 2007). N-acetyl-glucosamine (GlcNAc) and glucuronic acid (GlcUA) monomers polymerize via b1,3- and b-1,4-glycosidic bonds to form HA chains, the length of which determines the molecular weight (MW), and different HAs have different functions (Stern et al. 2006). High-molecular-weight (HMW) HAs (MW [ 2 MDa) have good viscoelasticity, strong rigidity, and maintain their properties for a long time. HMW HAs can be used in ophthalmic surgery and are useful for wound healing. Mediummolecular-weight (MMW) HAs (MW 0.1-1 MDa) display good moisture retention and lubricity, and they are suitable for soft tissue filling, deep wrinkle removal, and drug release. Low-molecular-weight
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(LMW) HAs (MW \ 10 kDa) have superior permeability and cell penetration properties, can inhibit tumor proliferation, promote bone formation and angiogenesis, and be used in immunomodulation (Stern et al. 2006; Toole et al. 2008). In recent years, heterologous HA-producing strains with clear backgrounds have become established as an attractive alternative for HA production due to the high pathogenic risk and paucity of DNA manipulation techniques for Stre
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