Comparison of two high-pressure homogenization methods for disrupting Hansenula polymorpha cells
Objective To compare the disrupting effects of Hansenula polymorpha cells by two kinds of high-pressure homoge-nization methods,step-by-step disrupting and cyclic disrupting,so as to provide experimental basis for the selection of disrupting methods in the industrial large-scale production of Hansenula polymorpha cells in the future.Methods Three batches of concentrated fermentation broth of Hansenula polymorpha cells were disrupted by two high-pressure homogenization methods:step-by-step disrupting and cyclic disrupting.The samples before and after disrupting were taken,and the cell morphology was observed under light microscope.The host protein content,cell disruption rate,protein release rate and particle size distribution of released protein were detected and compared.Results After disrupting three batches of concentrated fermentation broth of Hansenula polymorpha cells by step-by-step disrupting and cyclic disrupting,the number of intact cells decreased and cell fragments increased,while there was no obvious difference in cell morphology.In addition,there was no significant dif-ference in host protein content(average gray value of 115.259 and 109.125 respectively),cell disruption rate(average value of 60.29％and 58.96％respectively),protein release rate(average value of 39.55％and 37.28％respectively),and released protein particle size distribution(average size of 90.41 and 525.07 nm after step-by-step disrupting,accounting for 31.37％and 68.63％of sample protein content;average size of 109.11 and 547.37 nm after cyclic disrupting,accounting for 29.93％and 70.07％of sample protein content,respectively.F=1.054,1.159,4.245 and 8.875,t=1.036 0,0.504 1,0.605 0 and 0.139 7,respectively,each P＜0.05).Conclusion Cyclic and step-by-step high-pressure homogenization disrupting methods have the equivalent disrupting effect on Hansenula polymorpha cells,while cyclic disrupting is simpler and can avoid pollution risks,which is more suitable for the disrupting of Hansenula polymorpha cells for industrial large-scale production.
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