Kim Sung Jin, Yim Joung Han
Polar BioCenter, Korea Polar Research Institute, KORDI, Incheon, Republic of Korea.
J Microbiol. 2007 Dec;45(6):510-4.
Twenty-five bacterial strains that secrete mucous materials were isolated from sediment obtained from King George Island, Antarctica. Seven of these strains proved capable of producing cryoprotective exopolysaccharides. The strain KOPRI 21653 was selected for the further study of an anti-ice-nucleating polysaccharide (ANP), which originated from a polar region. KOPRI 21653 was identified as Pseudoalteromonas arctica as the result of 16S rRNA analysis. The exopolysaccharide, P-21653, was purified completely from the KOPRI 21653 cell culture via column chromatography and protease treatment. The principal sugar components of P-21653 were determined to be galactose and glucose, at a ratio of 1:1.5, via GC-MS analysis. The cryoprotective activity of P-21653 was characterized via an E. coli viability test. In the presence of 0.1% (w/v) P-21653, the survival ratio of E. coli cells was as high as 82.6% over three repeated freeze-thaw cycles. The survival ratio decreased drastically to 71.5 and 48.1%, respectively, in five and seven repeated cycle conditions; however, the survival ratios were greater over three (96.6-92.1%) to seven (100.5-91.6%) freeze-thaw cycles in the presence of 0.5 and 1.0% (w/v) P-21653. In addition, at much lower concentrations (0.1-1.0%), P-21653 resulted in survival ratios (83.1-98.4%) similar to those of two commercially available cryoprotectants (VEG plus X-1000, 92.9% and VM3, 95.3%), which were utilized at the recommended concentrations (90%). The biochemical characteristics of exopolysaccharide P-21653 reflect that this compound may be developed as a useful cryoprotectant for use in medical applications and in the food industry.
从南极乔治王岛采集的沉积物中分离出25株分泌黏液物质的细菌菌株。其中7株被证明能够产生具有冷冻保护作用的胞外多糖。选择菌株KOPRI 21653进一步研究源自极地地区的抗冰核多糖(ANP)。经16S rRNA分析,KOPRI 21653被鉴定为北极假交替单胞菌。通过柱色谱和蛋白酶处理从KOPRI 21653细胞培养物中完全纯化出胞外多糖P-21653。通过气相色谱-质谱分析确定P-21653的主要糖成分是半乳糖和葡萄糖,比例为1:1.5。通过大肠杆菌活力测试表征P-21653的冷冻保护活性。在存在0.1%(w/v)P-21653的情况下,经过三次重复冻融循环,大肠杆菌细胞的存活率高达82.6%。在五次和七次重复循环条件下,存活率分别急剧下降至71.5%和48.1%;然而,在存在0.5%和1.0%(w/v)P-21653的情况下,经过三次(96.6%-92.1%)至七次(100.5%-91.6%)冻融循环,存活率更高。此外,在低得多的浓度(0.1%-1.0%)下,P-21653产生的存活率(83.1%-98.4%)与两种市售冷冻保护剂(VEG plus X-1000,92.9%和VM3,95.3%)在推荐浓度(90%)下的存活率相似。胞外多糖P-21653的生化特性表明,这种化合物可能被开发为一种用于医疗应用和食品工业的有用冷冻保护剂。