CAJ | 학술논문

采用超高压技术处理胰蛋白酶，改变其空间结构，研究酶空间结构变化与酶活力之间的关系。采用傅立叶红外光谱（FTIR）检测超高压处理后胰蛋白酶的二级结构变化；采用荧光光谱检测处理后胰蛋白酶的三级结构；酶活力的检测采用福林酚法。结果显示，与未处理的相比，在37℃，不同压力（100～600 M Pa ）条件处理20 min ，对胰蛋白酶活力影响显著（p＜0.05）。其中，300 MPa处理，胰蛋白酶活力达到最大，较未处理的酶活提高了0.386倍。FTIR检测分析显示，300 MPa处理的胰蛋白酶，α-螺旋与β-转角的峰面积比值达到最大（2.749）；内源性荧光光谱检测结果显示，当激发波长为295 nm ，其荧光强度达到最高值（1353）；激发波长为280 nm ，其荧光强度达到最高（4262）；外源性荧光光谱结果显示，当激发波长为228 nm ，疏水氨基酸残基的荧光强度达到最高（2022）；上述荧光强度的变化较0.1 M Pa处理的胰蛋白酶均有显著差异（p＜0.05）。结论：超高压处理影响胰蛋白酶的空间结构及酶活性。其中，胰蛋白酶活性与α-螺旋和β-转角的峰面积的比值、色氨酸等疏水氨基酸及酪氨酸残基暴露程度有关。
채용초고압기술처리이단백매，개변기공간결구，연구매공간결구변화여매활력지간적관계。채용부립협홍외광보（FTIR）검측초고압처리후이단백매적이급결구변화；채용형광광보검측처리후이단백매적삼급결구；매활력적검측채용복림분법。결과현시，여미처리적상비，재37℃，불동압력（100～600 M Pa ）조건처리20 min ，대이단백매활력영향현저（p＜0.05）。기중，300 MPa처리，이단백매활력체도최대，교미처리적매활제고료0.386배。FTIR검측분석현시，300 MPa처리적이단백매，α-라선여β-전각적봉면적비치체도최대（2.749）；내원성형광광보검측결과현시，당격발파장위295 nm ，기형광강도체도최고치（1353）；격발파장위280 nm ，기형광강도체도최고（4262）；외원성형광광보결과현시，당격발파장위228 nm ，소수안기산잔기적형광강도체도최고（2022）；상술형광강도적변화교0.1 M Pa처리적이단백매균유현저차이（p＜0.05）。결론：초고압처리영향이단백매적공간결구급매활성。기중，이단백매활성여α-라선화β-전각적봉면적적비치、색안산등소수안기산급락안산잔기폭로정도유관。
Trypsin was treated by high pressure technology ,and its spatial structure was changed ,the relationship between structural changes and trypsin activity was investigated .The secondary structure change of trypsin after pressure treatment was observed by Fourier transform infrared spectroscopy (FTIR) .Moreover its tertiary structure change was observed by fluores-cence spectroscopy ;and its activity was tested using Folin phenol method .The results showed that ,compared with the untreat-ed(0.1 MPa) ,trypsin activity change was significant(p<0.05) under different pressure(100~600 MPa) treatment at 37 ℃ for 20 min .After treated with 300 MPa ,its activity was 0 .386 times higher than the untreated .Secondary structure of trypsin was analysed using FTIR ,and the peak area ratio of α-helix and β-turn in secondary structure was the maximum(2 .749);Endoge-nous fluorescence spectra intensity was the maximum (1 353) at excitation wavelength 295 nm ,and was 4 262 at excitation wavelength 280 nm ;exogenous fluorescent spectra intensity was 2 022 at excitation wavelength 228 nm ,all these change was re-markable(p<0.05)comparing with the untreated .Therefore ,ultrahigh pressure processing influence on the spatial structure of trypsin and induce enzyme activity change .T rypsin activity is relate to the peak area ratio of α-helix andβ-turn and the exposure degree of Trp and other hydrophobic a mino acid residues and Tyr .