College of Marine Life Sciences, Ocean University of China, Yushan Road, No. 5, Qingdao, China.
College of Marine Life Sciences, Ocean University of China, Yushan Road, No. 5, Qingdao, China; Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, 266003, China.
Int J Biol Macromol. 2019 Jun 1;130:988-996. doi: 10.1016/j.ijbiomac.2019.03.039. Epub 2019 Mar 6.
Aureobasidium melanogenum 11-1 was found to be able to produce over 281.7 ± 7.1 U/mL of β-fructofuranosidase activity. The protein deduced from the cloned β-fructofuranosidase1 gene had the conserved motif A (IGDP), motif D (RDP) and motif E (ET) and 11 N-glycosylation sites, indicating it was a β-fructofuranosidase with the high-level fructooligosaccharide (FOS) biosynthesis. Overexpression of the β-fructofuranosidase1 gene in the yeast strain 11-1 made a tranformant 33 produce 557.7 U/mL of β-fructofuranosidase activity. The molecular weight of the β-fructofuranosidase1 in which all the carbohydrates were removed by the Endo-H was 82.4 kDa. Within 7 h of the transfructosylation reaction, the yield of FOS was 0.66 g of FOS/g of sucrose and percentages of GF2, GF3 and GF4 were 79.5%, 18.9% and 1.6%. This demonstrated that the β-fructofuranosidase1 and the transformant 33 had highly potential applications in biotechnology for FOS production.
嗜鞣管囊酵母 11-1 被发现能够产生超过 281.7±7.1U/mL 的β-果聚糖酶活性。从克隆的β-果聚糖酶 1 基因推导的蛋白质具有保守基序 A（IGDP）、基序 D（RDP）和基序 E（ET）和 11 个 N-糖基化位点，表明它是一种具有高水平果寡糖（FOS）生物合成能力的β-果聚糖酶。在酵母菌株 11-1 中过表达β-果聚糖酶 1 基因，使转化体 33 产生 557.7U/mL 的β-果聚糖酶活性。经内切酶 H 去除所有碳水化合物后，β-果聚糖酶 1 的分子量为 82.4kDa。在转果糖反应的 7 小时内，FOS 的产率为 0.66g FOS/g 蔗糖，GF2、GF3 和 GF4 的百分比分别为 79.5%、18.9%和 1.6%。这表明β-果聚糖酶 1 和转化体 33 在生物技术中具有生产 FOS 的巨大潜力。
