Guo Lulu, Wang Shaolu, Lian Chunang, He Lisheng
Department of Deep-sea Science Research, Institute of Deep-Sea Science and Engineering, Chinese Academy of Sciences, Sanya, Hainan, China.
PeerJ. 2025 Apr 25;13:e19253. doi: 10.7717/peerj.19253. eCollection 2025.
Hyaluronan synthases (HASs) are ubiquitous in living organisms, and the hyaluronic acid (HA) synthesized by them are important to their body and well used in medicine, cosmetics and other fields. HAS from deep-sea creatures has not yet been explored before. The study aims to analyse the characteristics and enzyme kinetics of a novel hyaluronan synthase derived from the symbiont " Mycoplasma liparidae" found in deep-sea snailfish (snHAS).
snHAS was over-expressed using His as tag in the study. The sequence alignment was conducted by Cluster and then the phylogenetic analyse of HASs was performed by Mega 6.0 to investigate the position of snHAS during evolution. and were detected to study the enzyme kinetics of snHAS wildtype and its mutant. The molecular weight of HA was evaluated by high performance gel permeation chromatography (HPGPC). The cardiolipin was added to investigate whether it had a promoting effect on the snHAS.
The length of snHAS was 933 bp with an open reading frame (ORF) of 310 amino acids. Unlike other repoted HASs, snHAS had no transmembrane region and was not classified into the currently known Class I or Class II. snHAS could synthesize hyaluronan with lower molecular weights using the substrates of uridine-diphosphate--acetylglucosamine (UDP-GlcNAc) and uridine-diphosphate-glucuronic acid (UDP-GlcA) . The values of snHAS were 258 ± 45 µM and 39 ± 5 µM for UDP-GlcNAc and UDP-GlcA, respectively, much lower than those from mice ( for UDP-GlcA: 55 ± 5 µM; for UDP-GlcNAc: 870 ± 60 µM). The / values of snHAS were 163.5 s mM and 8.08 s mM for UDP-GlcA and UDP-GlcNAc, respectively. Furthermore, the activity of snHAS was independent of cardiolipin.
snHAS was a novel HAS based on the characteristics of the animo acid sequence, which could produce low molecular weight of HA with high efficiency. This provides a molecular basis for the biosynthesis of low molecular weight of HA.
透明质酸合酶（HASs）在生物体内广泛存在，由它们合成的透明质酸（HA）对生物体很重要，并在医学、化妆品和其他领域有广泛应用。此前尚未对深海生物来源的HAS进行过探索。本研究旨在分析一种源自深海狮子鱼共生菌“脂类支原体”的新型透明质酸合酶（snHAS）的特性和酶动力学。
在本研究中，以His为标签对snHAS进行了过表达。通过Cluster进行序列比对，然后使用Mega 6.0对HASs进行系统发育分析，以研究snHAS在进化过程中的位置。检测Km和Vmax以研究snHAS野生型及其突变体的酶动力学。通过高效凝胶渗透色谱法（HPGPC）评估HA的分子量。添加心磷脂以研究其对snHAS是否有促进作用。
snHAS的长度为933 bp，开放阅读框（ORF）为310个氨基酸。与其他已报道的HASs不同，snHAS没有跨膜区域，也未被归类到目前已知的I类或II类。snHAS可以使用尿苷二磷酸-N-乙酰葡糖胺（UDP-GlcNAc）和尿苷二磷酸葡糖醛酸（UDP-GlcA）作为底物合成分子量较低的透明质酸。snHAS对UDP-GlcNAc和UDP-GlcA的Km值分别为258±45μM和39±5μM，远低于小鼠来源的（UDP-GlcA的Km：55±5μM；UDP-GlcNAc的Km：870±60μM）。snHAS对UDP-GlcA和UDP-GlcNAc的Vmax/Km值分别为163.5 s mM和8.08 s mM。此外，表示snHAS的活性与心磷脂无关。
基于氨基酸序列特征，snHAS是一种新型的HAS，它可以高效地产生低分子量的HA。这为低分子量HA的生物合成提供了分子基础。
