Optimization of Teaching Experiment of Yeast Alcohol Dehydrogenase Extraction and Purification

粗提料液比	蛋白质总量/mg	总活力/U	比活力/（U·mg⁻¹）
注：同一因素同一测定指标不同水平间进行方差分析和多重比较，同一字母表示各水平间无显著差异，不同字母表示各水平间差异显著（P<0.05）。下同。
1:3	16.77±1.57^a	640.89±272.75^a	37.77±15.08^a
1:4	38.60±9.31^ab	2116.00±81.68^b	57.10±14.51^ab
1:5	48.85±6.10^b	4675.56±600.20^c	97.81±25.23^bc
1:6	79.54±5.40^c	10313.33±1524.74^d	130.03±21.01^c
1:7	120.24±24.07^d	11613.33±533.83^d	99.68±23.39^bc

Table 2. [in Chinese]

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Table 2. [in Chinese]

粗提温度/ ℃	蛋白质总量/mg	总活力/U	比活力/（U·mg⁻¹）
25	49.00±3.92^a	12120.00±432.67^c	247.94±11.30^c
35	43.79±5.89^a	8960.00±554.26^b	206.09±16.34^b
45	52.85±1.92^a	8537.78±398.01^b	161.72±10.28^a
55	44.88±2.71^a	6875.56±559.06^a	153.16±7.91^a
65	51.08±3.90^a	6624.44±690.78^a	130.63±20.87^a

Table 3. [in Chinese]

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Table 3. [in Chinese]

粗提时间/h	蛋白质总量/mg	总活力/U	比活力/（U·mg⁻¹）
1.5	21.17±3.55^a	2311.11±472.93^a	113.04±36.57^a
2.0	27.18±1.75^a	2113.33±396.76^a	78.62±20.21^a
2.5	24.79±5.31^a	2233.33±560.46^a	92.84±28.26^a
3.0	24.62±3.51^a	2423.11±567.47^a	97.76±13.93^a
3.5	22.12±4.97^a	1834.67±253.62^a	88.18±34.92^a

Table 4. [in Chinese]

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Table 4. [in Chinese]

热变性温度/ ℃	上清液蛋白质总量/mg	上清液总活力/U	上清液比活力/（U·mg⁻¹）	沉淀蛋白质总量/mg	沉淀总活力/U
35	49.97±1.56^d	10282.22±210.43^b	205.93±8.41^a	1.40±0.12^a	248.89±342.89^a
45	48.56±1.58^d	10342.22±1022.10^b	213.14±22.35^a	1.38±0.15^a	497.78±427.78^a
55	43.21±3.29^c	10735.56±957.32^b	248.38±8.85^a	5.50±0.08^b	355.56±222.04^a
65	21.89±0.77^b	12535.56±1196.30^c	571.74±34.03^c	7.77±0.32^c	106.67±106.67^a
75	12.80±0.92^a	6117.78±465.06^a	478.15±25.85^b	8.06±2.11^c	35.56±61.58^a

Table 5. [in Chinese]

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Table 5. [in Chinese]

热变性时间/min	上清液蛋白质总量/mg	上清液总活力/U	上清液比活力/（U·mg⁻¹）	沉淀蛋白质总量/mg	沉淀总活力/U
10	42.24±4.49^a	20533.33±2120.13^a	487.08±36.91^a	2.50±0.20^a	400.00±230.94^a
15	33.11±6.63^a	19724.44±1214.53^a	619.22±176.15^a	3.51±0.11^b	222.22±153.96^a
20	41.86±6.37^a	19920.00±739.01^a	482.03±63.07^a	3.59±0.18^b	266.67±230.94^a
25	41.06±14.43^a	20720.00±510.69^a	542.24±161.16^a	3.76±0.06^b	844.44±277.56^a
30	39.33±6.64^a	19160.00±1091.14^a	499.15±107.61^a	3.72±0.17^b	355.56±504.79^a

Table 6. [in Chinese]

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Table 6. [in Chinese]

有机溶剂种类	上清液蛋白质总量/mg	上清液总活力/U	上清液比活力/（U·mg⁻¹）	沉淀蛋白质总量/mg	沉淀总活力/U
乙醇	26.51±2.42^a	27002.22±2850.16^b	1026±166.54^a	7.79±0.29^b	17.78±30.79^a
丙酮	32.77±3.65^a	21528.89±1508.81^a	661.02±66.82^a	6.89±0.21^a	17.78±30.79^a
异丙醇	26.23±5.71^a	21073.33±697.07^a	832.36±201.41^a	8.30±0.25^c	17.78±30.79^a

Table 7. [in Chinese]

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Table 7. [in Chinese]

有机溶剂沉淀杂蛋白比例/倍	上清液蛋白质总量/mg	上清液总活力/U	上清液比活力/（U·mg⁻¹）	沉淀蛋白质总量/mg	沉淀总活力/U
0.3	73.89±9.09^b	19817.78±557.19^b	270.84±32.48^a	3.19±0.06^a	17.78±30.79^a
0.4	60.95±8.17^b	18777.78±529.54^b	311.34±36.51^a	3.02±0.05^a	17.78±30.79^a
0.5	60.36±6.45^b	20408.89±2363.63^b	337.88±5.19^a	3.62±0.17^b	0.00±0.00^a
0.6	41.71±4.20^a	20042.22±1132.83^b	485.42±74.61^b	3.97±0.09^c	853.33±488.81^b
0.7	31.27±4.73^a	16071.11±807.67^a	523.54±99.02^b	5.05±0.32^d	2133.33±774.71^c

Table 8. [in Chinese]

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Table 8. [in Chinese]

有机溶剂沉淀酶蛋白比例/倍	沉淀蛋白质总量/mg	沉淀总活力/U	沉淀比活力/（U·mg⁻¹）	上清液蛋白质总量/mg	上清液总活力/U
0.35	1.32±0.27^a	195.56±10.18^a	151.04±20.95^a	8.16±1.09^b	183.00±6.32^a
0.45	3.60±1.18^b	520.00±43.72^d	151.85±34.63^a	5.12±3.29^a	196.56±39.83^a
0.55	1.98±1.08^a	395.56±31.51^c	239.93±111.38^a	1.56±0.18^a	259.02±26.00^b
0.65	1.62±0.10^a	402.22±36.72^c	248.91±30.35^a	1.58±0.33^a	265.47±21.14^b
0.75	1.88±0.51^a	304.44±44.39^b	168.53±42.35^a	1.76±0.59^a	250.44±11.51^b

Table 9. [in Chinese]

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Table 9. [in Chinese]

提纯步骤	总体积/mL	蛋白质浓度 /（mg·m⁻¹）	蛋白质总量/mg	酶活力/U	总活力/U	比活力/（U·mg⁻¹）	回收率/%
提纯步骤	总体积/mL	蛋白质浓度 /（mg·m⁻¹）	蛋白质总量/mg	酶活力/U	总活力/U	比活力/（U·mg⁻¹）	蛋白质	酶活力
粗提	19.00	3.99	75.79	666.67	12666.67	167.12	100.00	100.00
热变性去除杂蛋白	17.50	1.68	29.36	846.67	14816.67	504.71	38.73	116.97
有机溶剂沉淀杂蛋白	24.00	0.73	17.52	613.33	14720.00	840.10	23.12	116.21
有机溶剂沉淀酶蛋白	5.00	0.90	4.52	826.67	4133.33	914.47	5.96	32.63

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Huajie GU, Tian MIU, Hengwei LIU, Hongwei CHEN, Weiqiang GUO, Taoyun WANG. Optimization of Teaching Experiment of Yeast Alcohol Dehydrogenase Extraction and Purification[J]. Experiment Science and Technology, 2025, 23(4): 25

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Paper Information

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Received: Feb. 14, 2024

Accepted: Mar. 31, 2025

Published Online: Jul. 30, 2025

The Author Email: Huajie GU (liongu@usts.edu.cn)

DOI:10.12179/1672-4550.20240050

Topics

laser devices and laser physics

Lasers and Laser Optics

Laser physics

laser manufacturing

Instrumentation, Measurement and Metrology