Department of Natural Science and Technology, Tay Nguyen University, Buon Ma Thuot 630000, Vietnam.
Department of Chemistry, Tamkang University, New Taipei City 25137, Taiwan.
Mar Drugs. 2020 Nov 20;18(11):574. doi: 10.3390/md18110574.
Microbial fermentation of by-products is a renewable and efficient technique in the development of a range of useful products. In this study, protease synthesis by sp. TKU052 was carried out on culture media containing some common seafood processing by-products (SPBPs) as the sole source of carbon and nitrogen (C/N). The most suitable C/N nutrition source for the production of proteases was found to be 3.0% () demineralized crab shells powder (deCSP) and maximal enzyme activity of 4.41 ± 0.16 U/mL was detected on the third day of the culture. Two proteases (P1 and P2) with a similar molecular weight of 31 kDa were successfully isolated and purified from the 3-day deCSP-containing medium. Both P1 and P2 exhibited the highest activity of gelatin hydrolysis at pH 6 and 60 °C. The gelatin hydrolysates catalyzed by TKU052 proteases were evaluated for biological activities, including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, angiotensin-I converting enzyme (ACE) inhibition, and prebiotic activities. The gelatin hydrolysates expressed 31.76-43.95% DPPH radical scavenging activity and 31.58-36.84% ACE inhibitory activity, which was higher than those from gelatin. Gelatin hydrolysates also showed the growth-enhancing effect on BCRC 14615 with an increase to 135.70-147.81%. In short, sp. TKU052 could be a potential strain to utilize crab shell wastes to produce proteases for bio-active peptides' preparation.
微生物发酵副产物是开发一系列有用产品的可再生且高效的技术。在这项研究中,利用一些常见的海鲜加工副产物(SPBPs)作为唯一的碳源和氮源(C/N),对 sp. TKU052 进行了蛋白酶合成。结果发现,最适合生产蛋白酶的 C/N 营养源是 3.0%(w/v)脱钙蟹壳粉(deCSP),在培养的第 3 天检测到最大酶活为 4.41 ± 0.16 U/mL。从含 3 天 deCSP 的培养基中成功分离和纯化了两种分子量相似的 31 kDa 的蛋白酶(P1 和 P2)。P1 和 P2 在 pH 6 和 60°C 时对明胶水解的活性最高。评估了 TKU052 蛋白酶催化的明胶水解产物的生物活性,包括 2,2-二苯基-1-苦基肼基(DPPH)自由基清除、血管紧张素转化酶(ACE)抑制和益生元活性。明胶水解产物的 DPPH 自由基清除活性为 31.76-43.95%,ACE 抑制活性为 31.58-36.84%,均高于明胶的相应活性。明胶水解产物对 BCRC 14615 的生长促进作用也有所提高,达到 135.70-147.81%。总之,sp. TKU052 可能是一种利用蟹壳废物生产生物活性肽制备用蛋白酶的潜在菌株。