蛋白質(zhì)提取分離分析及米氏常數(shù)測(cè)定的實(shí)踐及效果
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中圖分類號(hào) G642.1 文獻(xiàn)標(biāo)識(shí)碼A 文章編號(hào) 1007-7731（2025）13-0118-04
DOI號(hào)10.16377/j.cnki.issn1007-7731.2025.13.030
Practice and effect of protein extraction， separation， analysis， and Michaelis constant determination
JIA Haoran HAN Peng ZHANG Yantao ZHANG Liquan （CollegeofLife Sciences，Inner Mongolia University，HohhotO1oO21， China）
AbstractProtein extraction，separation，analysis and determination of Michaelis constant are the comprehensive experimental coursecontents inthe teaching sylabusofbiochemistryexperiments.In thisexperimental practice， commercially available mung bean sprout stem segments were used as experimental materials.The total protein was extracted bygrinding method，andtheconcentrationand distributionof total protein were determinedby BCA method and SDS-PAGE electrophoresis.The Michaelis constant of acid phosphatase in total protein was determined using disodium p-nitrophenyl phosphate （PNPP-N ）as the substrate under acidic conditions[37 °C ，0.2 mol/L HAc-NaAc α bufr （pH=5.6）.The results showed that the total protein concentration of mung bean sprout stem segments was 4.374 mg/mL.Under acidic conditions，the Km of acid phosphatase in the total protein was 5.552 5 mmol/L，and the Vm was 0.0743mmol/（L·min）.This practical exercise significantly enhancedstudents’operational understandingof fundamentalbiochemicaltechniques including protein extraction/purification，quantitative analysis，and determination of Michaelis constant.Furthermore，it provided hands-on experience with esential methodologies such asprotein isolationand SDS-PAGE electrophoresis， while efectively fostering scientific curiosityand research enthusiasm among participants.
Keywords Biochemistry; extraction and determination of protein; Michaelis constant; mung bean sprouts
蛋白質(zhì)是高校生物化學(xué)課程的核心教學(xué)內(nèi)容之一。（剩余5522字）