Hanyu Zhu1, Dongmei Liu1, Yuanyuan Wang1, Danfeng Ren1, Liesheng Zheng2, Liguo Chen2, Aimin Ma3,4. 1. College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China. 2. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China. 3. College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China. aiminma@mail.hzau.edu.cn. 4. Key Laboratory of Agro-Microbial Resources and Utilization, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, China. aiminma@mail.hzau.edu.cn.
Abstract
OBJECTIVES: 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. RESULTS: 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. CONCLUSION: Tremella fuciformis can be used as a cell factory to produce hydrophobin on a large scale for the food industry.
OBJECTIVES: 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. RESULTS: 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. CONCLUSION:Tremella fuciformis can be used as a cell factory to produce hydrophobin on a large scale for the food industry.