An Optimized Protocol for Crush Cell and Isolation of Protein from Scenedesmus sp. and Ammonia Toxicity Analysis

doi:10.13560/j.cnki.biotech.bull.1985.2015.11.014

Abstract

Abstract: Propose an optimized protocol for crush cell and isolation of protein from Scenedesmus sp. and study the ammonia toxicity of this algal with this protocol. Liquid nitrogen grinding and ultrasonic crushing method was used to disrupt the algae and acetone precipitation and TCA-acetone precipitation methods to extract and purify protein. Results showed that we found that the combination of liquid nitrogen grinding and TCA-acetone precipitation was effective and efficient as the cell crushed adequately and the protein sample was purified well and the protein amount and abundance was higher. The preliminary inquiry of ammonia toxicity to Scenedesmus sp. with two-dimensional gel electrophoresis(2-DE)technology found that 85 protein spots decreased and 25 protein spots increased as the algae cultured with ammonia compared with nitrate nitrogen. Those proteins have been identified as involved in photosynthesis, carbon dioxide fixation, glycolytic, protein synthesis and degradation of ubiquitinated protein, cell division and material transportation, and radical scavenging pathway.

Key words: 2-DE, Scenedesmus, crush cell, protein purification, ammonia toxicity

Jia Qikun, Liang Qing, Wu Houbo, Wang Guanghua, Wu Hualian, Li Tao, Wang Bing, Xiang Wenzhou. An Optimized Protocol for Crush Cell and Isolation of Protein from Scenedesmus sp. and Ammonia Toxicity Analysis[J]. Biotechnology Bulletin, 2015, 31(11): 125-130.

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