Strategies for Increasing Heterologous Protein Expression in Pichia pastoris

doi:10.13560/j.cnki.biotech.bull.1985.2023-0857

Abstract

Abstract:

The production of heterologous proteins via recombinant expression system is a focus of genetic engineering research. Because of its ease of genetic manipulation, high degree of heterologous protein secretion, and post-translational modification, Pichia pastoris, a methylotrophic yeast, has become an essential strain for protein production in industrial applications and is frequently employed in the manufacturing of enzymes. Although some heterologous proteins can be expressed in P. pastoris, the amount of expression is modest and has to be increased. Therefore, it is critical to investigate the principle and method of boosting the expression of heterologous proteins in P. Pastoris in order to lower industrial production costs and improve economic benefits. In this review, the optimizing strategies for improving heterologous protein expression in P. Pastoris in recent years are summarized from six perspectives: Gene level, transcription level, translation level, protein folding and secretion levels, cell resistance level, and fermentation technology, with the goal of demonstrating comprehensively how to improve the expression of heterologous protein in P. Pastoris expression system.

Key words: Pichia pastoris, recombinant expression, heterologous protein, optimization, enzyme

RUZHA Yelizhati, YANG Yu. Strategies for Increasing Heterologous Protein Expression in Pichia pastoris[J]. Biotechnology Bulletin, 2024, 40(3): 118-134.

Figures/Tables 10

References 108

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优化策略Optimizing strategy	外源蛋白Heterologous protein	优化效果Optimizing effect	发酵规模Fermentation scale/L	参考文献Reference
密码子优化	果胶酶	酶活提升1.19倍	-	[6]
	α-淀粉酶	酶活提升2.31倍	5	[8]
	α-淀粉酶	酶活提升2.62倍	50	[8]
	木聚糖酶	酶活提升1.44倍	5	[16]
	豆血红蛋白	酶活提升4.5倍	10	[17]
高拷贝筛选	甘露聚糖酶	酶活提升2.2倍	-	[10]
	L-天冬酰胺酶	酶活提升1.4倍	-	[11]
	壳聚糖酶	酶活提升1.61倍	50	[18]
	明胶	蛋白表达量提高1.31倍	140	[19]

优化策略Optimizing strategy	外源蛋白Heterologous protein	优化效果Optimizing effect	发酵规模Fermentation scale/L	参考文献Reference
密码子优化	果胶酶	酶活提升1.19倍	-	[6]
	α-淀粉酶	酶活提升2.31倍	5	[8]
	α-淀粉酶	酶活提升2.62倍	50	[8]
	木聚糖酶	酶活提升1.44倍	5	[16]
	豆血红蛋白	酶活提升4.5倍	10	[17]
高拷贝筛选	甘露聚糖酶	酶活提升2.2倍	-	[10]
	L-天冬酰胺酶	酶活提升1.4倍	-	[11]
	壳聚糖酶	酶活提升1.61倍	50	[18]
	明胶	蛋白表达量提高1.31倍	140	[19]

优化策略Optimizing strategy	外源蛋白Heterologous protein	优化效果Optimizing effect	发酵规模Fermentation scale/L	参考文献Reference
启动子的选择和使用	几丁质酶	蛋白表达量提高1.5倍	1.5	[30]
	葡萄糖醛酸酯酶	酶活提升3.5倍	14	[53]
	南极假丝酵母脂肪酶B	蛋白表达量提高3倍	-	[42]
终止子的选择和使用	羧基酯酶	蛋白表达量提高2.5倍	1	[44]
转录因子的过表达	水蛭透明质酸酶	酶活提升1.47倍	3	[45]
	植酸酶	酶活提升20%	-	[46]
	果胶酶	酶活提升35%	-	[46]
转录因子的去表达	人表皮生长因子受体-2单抗	蛋白表达量提高1.5倍	1	[48]
	-	解除甘油环境对P_AOX1的抑制	-	[50]
	-	挽救过氧化物酶体相关缺陷	-	[52]

优化策略Optimizing strategy	外源蛋白Heterologous protein	优化效果Optimizing effect	发酵规模Fermentation scale/L	参考文献Reference
启动子的选择和使用	几丁质酶	蛋白表达量提高1.5倍	1.5	[30]
	葡萄糖醛酸酯酶	酶活提升3.5倍	14	[53]
	南极假丝酵母脂肪酶B	蛋白表达量提高3倍	-	[42]
终止子的选择和使用	羧基酯酶	蛋白表达量提高2.5倍	1	[44]
转录因子的过表达	水蛭透明质酸酶	酶活提升1.47倍	3	[45]
	植酸酶	酶活提升20%	-	[46]
	果胶酶	酶活提升35%	-	[46]
转录因子的去表达	人表皮生长因子受体-2单抗	蛋白表达量提高1.5倍	1	[48]
	-	解除甘油环境对P_AOX1的抑制	-	[50]
	-	挽救过氧化物酶体相关缺陷	-	[52]

优化策略Optimizing strategy	外源蛋白Heterologous protein	优化效果Optimizing effect	发酵规模Fermentation scale	参考文献Reference
翻译起始因子的过表达	纳米抗体vHH	蛋白表达量提高3倍	1 L	[55]
核糖体生物合成因子的过表达	增强型绿色荧光蛋白	蛋白表达和细胞生物量均提高20%	250 mL	[58]
核糖体生物合成因子的过表达	植酸酶	酶活提升26%	250 mL	[58]
其他翻译相关元件的过表达	β-半乳糖苷酶	酶活提升2倍	-	[59]