School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China.
Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
Molecules. 2022 May 2;27(9):2897. doi: 10.3390/molecules27092897.
A high enzyme-yield strain sp. 298 was screened from marine bacteria harvested from the coastal water. The screening conditions were extensive, utilizing hyaluronic acid (HA)/chondroitin sulfate (CS) as the carbon source. A coding gene of the family 8 polysaccharide lyase (PL8) was cloned from the genome of sp. 298 and subjected to recombinant expression. The specific activity of the recombinase YsHyl8A was 11.19 U/mg, with an optimal reaction temperature of 40 °C and 50% of its specific activity remaining after thermal incubation at 30 °C for 1 h. In addition, its optimal reaction pH was 7.5, and while it was most stable at pH 6.0 in NaHPO-citric acid buffer, it remained highly stable at pH 6.0-11.0. Further, its enzymatic activity was increased five-fold with 0.1 M NaCl. YsHyl8A, as an endo-lyase, can degrade both HA and CS, producing disaccharide end-products. These properties suggested that YsHyl8A possessed both significant alkalophilic and cold-adapted features while being dependent on NaCl, likely resulting from its marine source. is a typical fish pathogen, with glycosaminoglycan lyase (GAG lyase) as a potential pathogenic factor, exhibiting strong hyaluronidase and chondroitinase activity. Further research on the pathogenic mechanism of GAG lyase may benefit the prevention and treatment of related diseases.
一株产酶能力较高的 sp. 298 菌从海洋细菌中筛选出来，这些细菌取自沿海水域。筛选条件非常广泛，利用透明质酸（HA）/硫酸软骨素（CS）作为碳源。从 sp. 298 的基因组中克隆了一个家族 8 多糖裂解酶（PL8）的编码基因，并进行了重组表达。重组酶 YsHyl8A 的比活为 11.19 U/mg，最适反应温度为 40°C，在 30°C 下孵育 1 小时后仍保留 50%的比活。此外，其最适反应 pH 为 7.5，在 NaHPO-柠檬酸缓冲液中最稳定 pH 为 6.0，在 pH 6.0-11.0 范围内仍保持高度稳定性。进一步研究发现，其酶活在 0.1 M NaCl 存在时增加了五倍。作为一种内切酶，YsHyl8A 可以降解 HA 和 CS，产生二糖末端产物。这些特性表明，YsHyl8A 具有显著的嗜碱性和耐冷性，同时依赖于 NaCl，这可能与其海洋来源有关。 是一种典型的鱼类病原体，糖胺聚糖裂解酶（GAG 裂解酶）是一种潜在的致病因子，具有很强的透明质酸酶和软骨素酶活性。进一步研究 GAG 裂解酶的致病机制可能有助于相关疾病的预防和治疗。
