Zhu Hanyu, Liu Dongmei, Wang Yuanyuan, Ren Danfeng, Zheng Liesheng, Chen Liguo, Ma Aimin
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
Biotechnol Lett. 2017 Aug;39(8):1167-1173. doi: 10.1007/s10529-017-2343-0. Epub 2017 May 3.
To obtain hydrophobin, a Class I hydrophobin gene, Po.hyd from Pleurotus ostreatus, was transformed into the yeast-like cells of Tremella fuciformis using Agrobacterium tumefaciens.
The hydrophobin Po.HYD from P. ostreatus was heterogeneously expressed by the yeast-like cells of T. fuciformis. Plasmids harboring the Po.hyd gene driven by endogenous glyceraldehyde-3-phosphate dehydrogenase promoter were transformed by A. tumefaciens. The integration and expression of the rPo.HYD in the T. fuciformis cells were confirmed by PCR, Southern blot, fluorescence microscopy and quantitative real-time PCR. SDS-PAGE demonstrated that the rPo.HYD was extracted with the expected MW of 14 kDa. The yield of purified rPo.HYD was 0.58 mg/g dry wt. The protein, with its ability to stabilize oil droplets, exhibited a better emulsifying activity than the typical food emulsifiers Tween 20 and sodium caseinate.
Tremella fuciformis can be used as a cell factory to produce hydrophobin on a large scale for the food industry.
为获得疏水蛋白，将来源于平菇的I类疏水蛋白基因Po.hyd，利用根癌农杆菌转化至银耳的类酵母细胞中。
平菇的疏水蛋白Po.HYD在银耳的类酵母细胞中实现了异源表达。携带由内源性甘油醛-3-磷酸脱氢酶启动子驱动的Po.hyd基因的质粒经根癌农杆菌转化。通过PCR、Southern杂交、荧光显微镜和定量实时PCR证实了rPo.HYD在银耳细胞中的整合和表达。SDS-PAGE表明提取的rPo.HYD分子量为预期的14 kDa。纯化的rPo.HYD产量为0.58 mg/g干重。该蛋白具有稳定油滴的能力，其乳化活性优于典型的食品乳化剂吐温20和酪蛋白酸钠。
银耳可作为细胞工厂，为食品工业大规模生产疏水蛋白。
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