糙米发芽过程中γ-氨基丁酸（GABA）转化富集技术研究
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摘要
本文以粳糙米为试材，研究了不同培养条件、培养液组分对糙米发芽过程中GABA转化量的影响，优化了富含GABA发芽糙米培养条件，初步研究了发芽糙米中GAD的酶学特性。
1、研究了浸泡培养下，不同温度、通气量和培养时间对发芽糙米的生理活性和GABA等物质含量的影响。结果表明：随着温度的升高，糙米的生长速率加快，GABA累积量增加。通气有利于发芽糙米生长，以1.0 L min~(-1)时发芽糙米中GABA累积量最高。培养时间72h内，GABA含量随培养时间的延长而增加。利用响应面法优化糙米培养条件，得出浸泡培养条件下发芽糙米高效转化GABA的最佳条件为：培养温度33.43℃、通气量1.28 L min~(-1)和培养时间47.6h。
2、研究了糙米在浸泡培养过程中加入谷氨酸钠、抗坏血酸以及谷氨酸钠加入时间对发芽糙米GABA转化量的影响。在单因子试验的基础上，进行了正交试验。结果表明：谷氨酸钠为1mg mL~(-1)，抗坏血酸为5mg mL~(-1)以及培养24h后加入谷氨酸钠的条件下发芽糙米中GABA富积量高达203.47μg g~(-1)，比对照高出2.73倍。
3、发芽糙米中GAD酶学性质的研究结果表明：最适反应pH为5.5-6.0，最适反应温度为40℃，以L-Glu为底物，双到数作图法得到Km为1.69 mM，Languir作图法得到Km为1.43mM。
4、进行了富含γ-氨基丁酸精白米制备工艺的研究。结果表明，固定发芽糙米中的GABA以热水处理为最好，在温度90℃和加水率1:1的条件下处理6min，转入50℃干燥箱内干燥至安全含水量。对高效转化GABA发芽糙米进行碾白处理可有效地改善产品的感官质量，但造成产品的营养损失，碾白90s后得到的精白米中GABA含量为8.962mg(100g~(-1)DW)。
Effects of different cultivation conditions, composition of culture liquid onγ-aminobutyric acid (GABA) enrichment in geminating brown rice {Oryza stativa L. subsp. Japonaca) were investigated and process parameters for brown rice germination were optimized. The properties of L-glutamate decarboxylase (GAD) were preliminarily studied. The processing technology of fine rice rich in GABA was developed, which provide a guideline for germinated brown rice industrialization. In this dissertation, major working and findings were summarized as follows:1. Under soaking cultivation condition, effects of different temperatures, aerating volume and duration on the physiological activities of and content of GABA in germinating brown rice was investigated using single factor trial and Box-Behnken design. The growth rate of brown rice sprout increased and accumulation of GABA was accelerated with increase of temperature. Aerating was in favor of the growth of brown rice sprout. The maximum content of GABA in germinated brown rice was reached at an aerated volume of 1.0 L min-1. GABA content increased over time within 72 hours.The maximum GABA content in germinating brown rice had been observed at a speed of 1.28 L min~(-1) and temperature of 41.08℃germinated for 47.6 hours with response surface methodology.2. The brown rice was immersed into the solution with different additives concentration of monosodium glutamate (MSG) and ascorbic acid (ASA) for germination under the conditions of 32℃, aerated volume of 1.2 liter per minute and darkness. The influence of MSG, ASA and the time of MSG addition on the accumulation of GABA in germinating brown rice was studied with single and orthogonal trial designs. The results showed that under the optimum combination condition of 1 mg mL~(-1) MSG, 5 mg mL~(-1) ASA and after adding the MSG for 24 hours germination, the content of GABA in germinated brown rice reached 203.47μ.g g~(-1), which was 3.73 times higher than that in control group.3. GAD in germinated brown rice exhibited an optimum pH value range from 5.5 to 6.0 and optimum reaction temperature of 40℃. Analysis of Lineweaver-Burkand Langmuir for GAD with L-Glu substrate was carded out, the Km were 1.43 mM.On the other hand, the Km of 1.69 mM was calculated by the method of double - reciprocal plotting.
4. Investigation on the processing technology of fine rice rich in GABA was carried out. Germinated brown rice was subjected to blanching in hot water for inaction of intrinsic enzyme. The minimum GABA loss was obtained at the temperature of 90℃and 1:1 the ratio of rice to water for 6 minutes and then drying to safe water content at 50℃. Sensory quality of germinated brown rice rich in GABA could be improved through slight milling. But nutritional components might be loss during milling. The content of GABA in fine rice was 8.962 mg/100gDW after milling for 90 seconds, which was higher than normal fine rice.
1、研究了浸泡培养下，不同温度、通气量和培养时间对发芽糙米的生理活性和GABA等物质含量的影响。结果表明：随着温度的升高，糙米的生长速率加快，GABA累积量增加。通气有利于发芽糙米生长，以1.0 L min~(-1)时发芽糙米中GABA累积量最高。培养时间72h内，GABA含量随培养时间的延长而增加。利用响应面法优化糙米培养条件，得出浸泡培养条件下发芽糙米高效转化GABA的最佳条件为：培养温度33.43℃、通气量1.28 L min~(-1)和培养时间47.6h。
2、研究了糙米在浸泡培养过程中加入谷氨酸钠、抗坏血酸以及谷氨酸钠加入时间对发芽糙米GABA转化量的影响。在单因子试验的基础上，进行了正交试验。结果表明：谷氨酸钠为1mg mL~(-1)，抗坏血酸为5mg mL~(-1)以及培养24h后加入谷氨酸钠的条件下发芽糙米中GABA富积量高达203.47μg g~(-1)，比对照高出2.73倍。
3、发芽糙米中GAD酶学性质的研究结果表明：最适反应pH为5.5-6.0，最适反应温度为40℃，以L-Glu为底物，双到数作图法得到Km为1.69 mM，Languir作图法得到Km为1.43mM。
4、进行了富含γ-氨基丁酸精白米制备工艺的研究。结果表明，固定发芽糙米中的GABA以热水处理为最好，在温度90℃和加水率1:1的条件下处理6min，转入50℃干燥箱内干燥至安全含水量。对高效转化GABA发芽糙米进行碾白处理可有效地改善产品的感官质量，但造成产品的营养损失，碾白90s后得到的精白米中GABA含量为8.962mg(100g~(-1)DW)。
Effects of different cultivation conditions, composition of culture liquid onγ-aminobutyric acid (GABA) enrichment in geminating brown rice {Oryza stativa L. subsp. Japonaca) were investigated and process parameters for brown rice germination were optimized. The properties of L-glutamate decarboxylase (GAD) were preliminarily studied. The processing technology of fine rice rich in GABA was developed, which provide a guideline for germinated brown rice industrialization. In this dissertation, major working and findings were summarized as follows:1. Under soaking cultivation condition, effects of different temperatures, aerating volume and duration on the physiological activities of and content of GABA in germinating brown rice was investigated using single factor trial and Box-Behnken design. The growth rate of brown rice sprout increased and accumulation of GABA was accelerated with increase of temperature. Aerating was in favor of the growth of brown rice sprout. The maximum content of GABA in germinated brown rice was reached at an aerated volume of 1.0 L min-1. GABA content increased over time within 72 hours.The maximum GABA content in germinating brown rice had been observed at a speed of 1.28 L min~(-1) and temperature of 41.08℃germinated for 47.6 hours with response surface methodology.2. The brown rice was immersed into the solution with different additives concentration of monosodium glutamate (MSG) and ascorbic acid (ASA) for germination under the conditions of 32℃, aerated volume of 1.2 liter per minute and darkness. The influence of MSG, ASA and the time of MSG addition on the accumulation of GABA in germinating brown rice was studied with single and orthogonal trial designs. The results showed that under the optimum combination condition of 1 mg mL~(-1) MSG, 5 mg mL~(-1) ASA and after adding the MSG for 24 hours germination, the content of GABA in germinated brown rice reached 203.47μ.g g~(-1), which was 3.73 times higher than that in control group.3. GAD in germinated brown rice exhibited an optimum pH value range from 5.5 to 6.0 and optimum reaction temperature of 40℃. Analysis of Lineweaver-Burkand Langmuir for GAD with L-Glu substrate was carded out, the Km were 1.43 mM.On the other hand, the Km of 1.69 mM was calculated by the method of double - reciprocal plotting.
4. Investigation on the processing technology of fine rice rich in GABA was carried out. Germinated brown rice was subjected to blanching in hot water for inaction of intrinsic enzyme. The minimum GABA loss was obtained at the temperature of 90℃and 1:1 the ratio of rice to water for 6 minutes and then drying to safe water content at 50℃. Sensory quality of germinated brown rice rich in GABA could be improved through slight milling. But nutritional components might be loss during milling. The content of GABA in fine rice was 8.962 mg/100gDW after milling for 90 seconds, which was higher than normal fine rice.
引文
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