Lin Bokun, Liu Yan, Lu Guoyong, Zhao Min, Hu Zhong
Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou, Guangdong 515063, PR China.
Dongguan Key Laboratory of Environmental Medicine, School of Public Health, Guangdong Medical University, Dongguan, Guangdong 515063, PR China.
FEMS Microbiol Lett. 2017 Feb 1;364(4). doi: 10.1093/femsle/fnx012.
A novel β-agarase gene aga672 was cloned from strain ZC1, the typical strain of agar-degrading Aquimarina agarilytica. Gene aga672 is composed of 2130 bp, and the encoded protein Aga672 showed an amino acid sequence identity of only 42% with reported agarases. Aga672 should belong to glycoside hydrolase family 16 according to the protein sequence similarity. The molecular mass of the recombinant Aga672 was estimated to be 98 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Aga672 decomposed agarose to produce neoagarotetraose, neoagarohexaose and neoagarooctaose as the main products. That is the main difference between Aga672 and other reported agarases of family GH16. The Km and Vmax for agarose degradation were 59.8 mg mL-1 and 154.3 U mg-1, respectively. The activity of Aga672 was stable at temperatures below 40°C and at pH 7.0-11.0 with the maximal agarase activity at 25°C and pH 7.0. The results showed that agarase Aga672 could be suitable to hydrolyze the gelated agarose. Thus, it has potential applications in the production of neoagarooligosaccharides directly from red alga.
从琼胶降解菌海栖琼胶杆菌(Aquimarina agarilytica)的典型菌株ZC1中克隆出一个新的β-琼胶酶基因aga672。基因aga672由2130个碱基对组成,其编码的蛋白Aga672与已报道的琼胶酶氨基酸序列同一性仅为42%。根据蛋白序列相似性,Aga672应属于糖苷水解酶家族16。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳估计重组Aga672的分子量为98 kDa。Aga672分解琼脂糖产生新琼脂四糖、新琼脂六糖和新琼脂八糖作为主要产物。这是Aga672与其他已报道的GH16家族琼胶酶的主要区别。琼脂糖降解的Km和Vmax分别为59.8 mg mL-1和154.3 U mg-1。Aga672的活性在40°C以下及pH 7.0 - 11.0时稳定,在25°C和pH 7.0时具有最大琼胶酶活性。结果表明,琼胶酶Aga672适合水解胶凝的琼脂糖。因此,它在直接从红藻生产新琼寡糖方面具有潜在应用。