Compare extracellular proteinase activity of three phynotypes of Penicillium marneffei

Abstract

Abstract: Objective To find the effective way to induce Penicillium marneffei (PM) to secrete extracellular proteinase and compare the extracellular proteolytic activity of three phynotypes of PM.Methods PM phynotypes (M/M and M/Y) and their parent strain B-6323 (P) were respectively inoculated with same amount into separate yeast carbon base (YCB) agar plates containing bovine hemoglobin (BHG) as the proteolytic substrate which had been adjusted to pH 4.0, pH 5.6, pH 7.2, and incubated at 25℃ and 37℃ for 6 days. The plates were stained with Coomassie blue G-250 to be observed clearly. Clear zones surrounding each clone result from broken down hemoglobin were measured to ascertain whether YCB-BHG agar can induce PM producing proteinase successfully, and to determine the best suitable condition for PM proteinase induced and compared the proteinase activity produced by the three isolates.Results YCB-BHG agar can successfully induce PM producing proteinase. Any three phynotypes of PM inoculated into medium at any three value of pH, the diameter of clear zones surrounded each clone increased greatly at 37℃ compared with at 25℃. It had statistically significant difference by t-test; The variance analysis showed that the diameters of clear zones surrounding each clone inoculated at pH4.0, 5.6, 7.2 medium at 37℃ had statistically significant difference (F-Values 97.198,P=0.0001). Student-Newman-Keuls (SNK) method was used to compare one to another. The diameters of clear zones surrounded the clone inoculated at pH4.0, pH5.6 medium and the clone inoculated at pH7.2 medium had statistically significant difference; No matter the culture temperature was 37℃ or 25℃, in any pH medium group, the M/M strain had greatest mean diameter of clear zone, then P strain, M/Y strain was lowest.Conclusions Extracellular proteinase of Penicillium marneffei growed at 37℃ in acidic medium (pH4.0 or pH5.6) showed greater proteolytic activity. The proteolytic activity produced by different morphotypes of Penicillium marneffei was different. The extracellular proteolytic activity of M/M strain and P strain was greater than M/Y strain after growth on the same condition.

Key words: Penicillium marneffei, extracellular proteinase, activity

CLC Number:

R379.9

WANG Peng, RAN Yu-ping, YIN Bin, ZHUANG Kai-wen, LIN Xin-yu, DAI Ya-ling. Compare extracellular proteinase activity of three phynotypes of Penicillium marneffei[J]. Chinese Journal of Mycology, 2014, 9(4): 193-198.

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