发芽糙米营养特性、γ-氨基丁酸富集及生理功效的研究
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摘要
稻谷砻谷后的糙米虽然营养丰富，但蒸煮性和适口性等较差，且脂肪含量高，长期贮藏易酸败而导致品质下降。糙米加工精米的过程中造成了大量的营养损失。因此，在传统稻谷加工基础上，结合现代生物技术对糙米进行深加工是今后稻谷及其制品深加工的主要发展趋势之一。
发芽糙米是糙米发芽至适当长的芽体，主要由幼芽和带种皮的胚乳构成，富含许多生物活性物质如γ-氨基丁酸(gamma aminobutyric acid，GABA)，是一种具有全营养概念、特殊营养价值的功能性主食。本研究在糙米发芽后组织化学、理化特性的变化以及γ-氨基丁酸(GABA)富集探索基础上，进一步探讨GABA对小鼠体内糖类、脂类和蛋白质代谢的影响。旨在从宏观和微观揭示糙米发芽过程中发生的变化，评价发芽糙米的营养价值；筛选富含GABA发芽糙米制备的工艺条件；开发富含GABA发芽糙米及其制品。
1糙米发芽过程中营养成分及抗营养因子、碳水化合物组成及其相关酶活性的变化
目的：研究不同品种糙米发芽过程中营养成分和植酸含量、碳水化合物组成及相关酶活性的变化。方法：Ⅱ优838、川香优2号、江镇粳糯和香粳99糙米分别在35℃下发芽60h，每间隔12h取样并进行化学分析。结果：不同品种糙米发芽过程中淀粉、直链淀粉、支链淀粉和抗营养因子植酸含量均呈下降趋势，其他营养成分先上升后下降。总淀粉酶活力、α-淀粉酶活力先增加后降低。4个品种糙米发芽24h时抗坏血酸含量最高；赖氨酸含量约是发芽前的3倍，除江镇粳糯在发芽24h最高外，其余3个品种均在发芽36h达到峰值。色氨酸含量除Ⅱ优838在发芽48h最高外，其余3个品种在发芽36h达到峰值，约是发芽前的2倍。Ⅱ优838和川香优2号糙米可溶性蛋白、游离氨基酸和还原糖的含量在发芽36h达到峰值，江镇粳糯和香粳99的峰值则出现在发芽48h。其中，籼稻糙米(Ⅱ优838和川香优2号)糙米发芽36h时，还原糖和可溶性糖含量、总淀粉酶和α-淀粉酶活力达到高峰，而粳稻糙米(香粳99)和糯稻糙米(江镇粳糯)高峰值则出现在发芽48h时。结论：发芽可提高糙米营养成分的含量，降低抗营养因子植酸的含量；并通过影响糙米淀粉酶的活性，改善碳水化合物的组成，有利于糙米品质的改善。
2发芽对糙米蛋白质及氨基酸组成特性的影响
目的：研究发芽过程中糙米蛋白质和氨基酸含量和组成特性的变化，并与精米和糙米比较，评价发芽糙米蛋白质的营养价值。方法：糙米在35℃下发芽48h，每隔6h取样并进行化学分析。结果：发芽可以增加糙米蛋白质、总氨基酸和必需氨基酸的含量，必需氨基酸指数提高。发芽过程中清蛋白和谷蛋白含量均呈上升趋势，球蛋白含量呈下降的趋势。除发芽24h外，醇溶蛋白的含量在整个发芽期间也呈现增加趋势，增加的蛋白质主要是谷蛋白。各发芽阶段糙米的必需氨基酸和总氨基酸的比值均大于55％。18种氨基酸中有12种的含量在发芽24h最高。发芽后赖氨酸和苏氨酸的含量增加较明显，但仍是限制性氨基酸；丝氨酸、谷氨酸、精氨酸和组氨酸含量均有所增加。发芽24h时必需氨基酸指数最大，与FAO／WHO模式相比，必需氨基酸组成模式更加合理。结论：发芽可以提高糙米蛋白质的营养价值，改善籼稻米的品质，其中以发芽24h为宜。
3发芽对糙米淀粉理化特性和超微结构的影响
目的：研究发芽对糙米淀粉理化特性和超微结构的影响。方法：以未发芽、发芽12h、24h、36h的糙米为原料提取淀粉，并对淀粉及其组分含量、淀粉糊的透明度、凝沉性质、冻融稳定性、酶解率以及粘度等进行分析。结果：糙米发芽后淀粉及其组分的含量、淀粉糊的酶解率以及淀粉糊的粘度均降低；淀粉糊的透明度和冻融稳定性均增强；凝沉稳定性、糊化温度和淀粉颗粒超微结构变化不明显。结论：发芽对糙米淀粉的理化特性具有一定的改善作用。其中发芽24h的糙米淀粉的理化特性变化最明显。
4糙米发芽过程中GABA富集的研究
目的：研究不同浸泡剂对糙米发芽过程中γ-氨基丁酸富集的影响。方法：采用蒸馏水、CaCl_2溶液和壳聚糖溶液浸泡糙米，并通过控制不同的发芽温度(A：25℃、35℃、45℃)、发芽时间(B：12h、24h、36h)和pH(C：5.6、7.0、8.4)的正交试验设计[L_9(3~4)]，研究γ-氨基丁酸富集的适宜条件。结果：(1)蒸馏水浸泡的糙米在发芽24h前其GABA的含量呈现增加的趋势，发芽24h达到高峰，然后GABA含量呈下降趋势；各发芽阶段糙米中GABA含量均高于糙米和精米。(2)随着发芽时间的延长，0.1％CaCl_2溶液浸泡的糙米中GABA含量呈逐渐增加趋势，而0.5％CaCl_2溶液浸泡的糙米中GABA含量先增加后降低转而又增加，两种浓度下发芽42h时GABA均达到峰值，但低浓度下GABA的峰值高于高浓度。(3)随着发芽时间的延长，0.1％、0.3％和0.5％壳聚糖浸泡后糙米发芽24h内GABA含量呈逐渐增加趋势，24h后GABA含量逐渐减少；3种浓度下糙米发芽24h时GABA均达到峰值，在整个发芽期间，壳聚糖浓度高低与糙米中GABA含量呈正相关。(4)三因素三水平正交设计试验结果表明，pH为5.6时GABA富集效果明显。各因素水平对GABA含量的影响从主到次的顺序为C＞A＞B。结合前面系列实验结果综合考虑，选择各因素的最佳水平组合为A_3B_2C_1，即发芽温度为45℃、发芽时间为24h、发芽pH为5.6。
结论：蒸馏水、低浓度的CaCl_2和壳聚糖浸泡均有利于糙米发芽过程GABA的富集；pH值是影响发芽糙米GABA富集的主要因素，其中酸性环境有利于GABA的富集，其次是发芽温度，再次是发芽时间。
5小鼠体内GABA生理功效的研究
目的：通过分析GABA对小鼠体内血清、肌肉和肝脏中与糖类、脂类和蛋白质等代谢相关的生理生化指标的影响，探讨小鼠体内GABA的生理功效。方法：56只体重22g左右的4w雄性昆明种小鼠(p＞0.05)，随机分成4组，每组设2个重复，每个重复7只，分别为CK组(对照组，GABA量为零)、A组(低剂量组，GABA量为12.5mg／L)、B组(中剂量组，GABA量为125mg／L)和C组(高剂量组，GABA量为1250mg／L)。饲养2d后每天早晨0.2mL／只空腹灌胃，连续18d；每周末清晨空腹称重一次；屠宰前禁食12h，眼球取血，分离血清；并迅速处死，取其肝脏和肌肉，分析肝脏和肌肉中蛋白质、脂肪和胆固醇的沉积以及血清生理生化指标。结果：(1)每周末各GABA组小鼠的平均体重均高于CK组(p＞0.05)；C组小鼠的平均体重最大，但与其他各组间差异不显著(p＞0.05)。(2)与CK组相比，各GABA组小鼠肝脏和肌肉中蛋白质含量均增加，以B组肝脏和肌肉中蛋白质含量最高，但各组小鼠肌肉中蛋白质的沉积差异不显著(p＞0.05)；而B组肝脏中蛋白质的沉积显著高于CK组(p＜0.01)、A组(p＜0.05)和C组(p＜0.05)。(3)与CK组相比，各GABA组小鼠肝脏和肌肉中胆固醇含量均下降，但各组间差异不显著(p＞0.05)。其中肝脏胆固醇以B组最低，而肌肉中以C组最低。各GABA组小鼠肝脏脂肪含量也均有所降低，但各组间差异不显著(p＞0.05)，其中以B组最低，比对照组降低2.25个百分点；而肌肉中脂肪的沉积却加强，其中B组沉积量最大，高于CK组(p＜0.01)、A组(p＜0.05)和C组(p＜0.05)。(4)与CK组相比，各GABA组小鼠血清中TP、ALB、GLOB、UA、TG、CHOL、HDL-C、LDL-C含量以及AKP、CHE、LDH、ALT的活性均增加。其中B组TP最高，与CK组间差异极显著(p＜0.01)，A组和C组与CK组间差异显著(p＜0.05)，各GABA组间差异不显著性(p＞0.05)。ALB以B组最高，各GABA组间无显著性差异(p＞0.05)，但均显著高于CK组(p＜0.05)。各组的GLOB、UA、TG、HDL-C的含量以及CK、CHE、LDH、ALT的活性间差异不显著(p＞0.05)，其中B组的GLOB、HDL-C和CK最高，A组的UA、CHE、LDH、ALT最高，TG以C组最高。各GABA组小鼠的A／G、GLU均比CK组有所下降，但各组间差异不显著(p＞0.05)，其中A组的GLU最低，B组的A／G最低。各GABA组的BUN、CREA、BUN／CR与CK组相比增减不一，其中C组的这三个指标均高于CK组(p＜0.05；p＞0.05；p＞0.05)，而A组和B组的均低于CK组(p＞0.05；p＞0.05；p＞0.05)，此外，C组的BUN也高于A组(p＜0.05)和B组(p＜0.05)。C组(p＜0.01)、A组(p＜0.05)和B组(p＜0.05)的CHOL均高于CK组，但A、B、C组间无显著性差异(p＞0.05)。与CK组相比，C组(p＜0.01)、B组(p＜0.05)和A组(p＞0.05)的LDL-C均增加，A组(p＞0.05)、C组(p＞0.05)和B组(p＜0.05)的AKP也增加。结论：GABA可促进小鼠生长；加强肌肉和肝脏中蛋白质的合成与沉积，减少胆固醇的沉积，降低肝脂含量，加强肌脂的沉积。
Brown rice dehulled from paddy has poor taste, lower cooking capability and high fat content though enriched with nutrition that, however, tends to decay because of acidification for long term stocking. Substantial nutrition loss will happen during the process from brown rice to polished one. Thus it will be the trend that conventional rice processing techniques combine with modern biological farther processing methods.
Germinated brown rice consists of brown rice with proper amount of sprout and endosperm with seed coat, which is enriched with many biological active components such as gamma aminobutyric acid (GABA) and thus regarded as functional food that combines broad spectrum nutrition with special nutrient. With the study of histochemical change, physicochemical property, and GABA accumulation during germinating of brown rice, the effect is farther investigated of GABA on the metabolism of carbohydrate, lipid and protein of mice. This investigation was aimed to reveal microcosmic and macroscopic variation of brown rice during germination, evaluate its nutrition, determine the optimum germinating condition and develop germinated brown rice products enriched with GABA.
1 Investigation of brown rice germination on nutrients, anti-nutrition factor, carbohydrate and related enzyme
Purpose: Investigation of germination on nutrients, phytic acid, carbohydrate content and related enzyme activity in brown rice. Method: Four types of brown rice (Ⅱyou838, Chuanxiangyou2, Xiangjing99, Jiangzhenjingnuo) were selected to germinate for 60 hours under 35℃. Samples were taken for each 1:2 hours and tested chemically. Result: For all types, tests revealed that the contents of starch, amylose, amylopectin and anti-nutrition factor phytic acid reduced, while other nutrition content, activity of total amylase andα-amylase increased at first and then reduced. For all four types, ascorbic acid contents reached the highest level at 24h, and the lysine contents were about 3 times higher than those before germination and reached the highest level at 36h except for Jiangzhenjingnuo whose apex was at 24 hours. Tryptophan content reached the highest level at 36h-about 2 times of original except forⅡyou838 whose apex was at 48h. The contents of soluble protein, free amino acid and reduced sugar inⅡyou838 and Chuanxiangyou2 reached the highest level at germination of 36h, while the highest values happened at germination of 48h for Xiangjing99 and Jiangzhenjingnuo. For indica type rice among them (Ⅱyou838 and Chuanxiangyou2), the max. contents of reduced and soluble sugar, total amylase and a-amylase activity happened at germinated 36h, while 48h for nonglutinous rice (Xiangjing99) and glutinous rice (Jiangzhenjingnuo). Conclusion: By germinating, the contents of nutrition increased and phytic acid as anti-nutrition factor reduced. Besides, carbohydrate composition and the quality of brown rice were improved through affecting the amylase activity.
2 Effect of germination on protein and amino acid composition properties of brown rice
Purpose: Assessing the protein nutrition value of germinated brown rice by investigating the content and composition variation of protein and amino acid during germination, comparing with brown rice and polished rice as reference. Method: Brown rice was kept at 35℃for 48 hours and samples were taken every 6h and tested chemically. Result: Germination can increase the content of crude protein (CP), total amino acid (TAA), essential amino acid (EAA) and its index (EAAI). Content of albumin and glutelin increased as well while dropping of globulin content during germination. Except germinating for 24 hours, the content of prolamine rose and mostly was in glutelin. During the process, the ratio of essential amino acid to total amino acid kept over 55%. 12 of 18 types of amino acid reached the highest level at germination of 24 hours. The content of lysine and the threonine increased substantially also after germination though as limited amino acid yet. The content of serine, glutamic acid, arginine and histidine all increased then. EAAI summited at germination of 24 hours and with had more preferable EAA composition compared with FAO/WHO. Conclusion: The nutritional value of protein and the quality of indica brown rice can be improved by germinating and 24h is the most preferred germinating time.
3 Effect of germination on starch physicochemieal properties and granular micro-structure in brown rice
Purpose: Investigation of the effect of germination on starch physicochemical property and granular micro-structure of brown rice. Method: Extracting starch from brown rice germinated for Oh, 12h, 24h and 36h and testing its starch content, components, paste clarity, retrogradation, freeze-thaw stability, enzymatic hydrolyzation and viscosity. Result: After germinating of brown rice, the content of starch and its component, enzymatic hydrolyzation and viscosity declined, whereas clarity and freeze-thaw stability enhanced. Paste retrogradation, gelatinization temperature and granular micro-structure of starch had no obvious change. Conclusion: Germinating resulted in improvement of physicochemical properties of brown rice starch and that at 24h was most significant.
4 Accumulation in brown rice of GABA during the germination
Purpose: Investigation of the effect of different soaking agents on the accumulation of gamma aminobutyric acid in germinated brown rice. Method: Soaking brown rice with distilled water, CaCl_2 solution and chitosan at different temperature (A: 25℃, 35℃, 45℃), time (B: 12h, 24h, 36h) and pH (C: 5.6, 7.0, 8.4) that was designed as three-factor and three-level orthogonal experiment [L_9 (3_4)] to determined the content variation of GABA in the process of brown rice germination. Result: (1) Within 24h, the GABA content in brown rice soaked in distilled water increased, then decreased and kept higher than that of brown rice and polish rice in every germination stage. (2) Along with the germination time, the GABA content of brown rice soaked in 0.1%CaCl_2 solution gradually increased, while that in 0.5%CaCl_2 solution increased at first and then reduced and increased again. The peak of GABA all happened at germination of 42h, and was higher in low concentration than that in high one. (3) Along with the germination time, the GABA content of brown rice soaked in 0.1%、0.3%、0.5% chitosan solution gradually increased within 24h then decreased. The peaks of GABA all happened at germination 24h. The content of GABA was positively correlated with that of chitosan. (4) The orthogonal experiment revealed that GABA accumulated substantially at pH5.6. Factors affecting GABA content ranked as C＞A＞B. Considering results of all series as a whole, the optimum condition of GABA accumulation in the process of brown rice germination was A_3B_2C_1, namely 45℃, 24h and pH5.6. Conclusion: Distilled water, low concentration CaCl_2 solution and chitosan all can enhance GABA accumulation of brown rice in germination, pH is the key factor affecting GABA accumulation, while second is germination temperature and time the third. Acid condition can promote GABA accumulation.
5 Physiological functions of GABA on mice
Purpose: Investigation of physiological functions of GABA on mice by analyzing the effects on biochemical parameters correlative to the metabolism of carbohydrate, lipid and protein in mice serum, muscle and liver. Method: 56 male Kunming mice of 4-w-old and weighing about 22g (p＞0.05) were randomly divided into 4 groups, and each group consisted of 14, and 7 of per group serving for one repeat. The groups were named as group CK (Control group, 0mg/L GABA), group A (low dose, 12.5mg/L GABA), group B (middle dose, 125mg/L GABA) and group C (high dose, 1250mg/L GABA). After two day's breeding, each mouse was filled in stomach with 0.2mL GABA for 18d at limosis state every morning, and was weighed every weekend morning under limosis state. With 12-hour fasting before slaughter, the blood sample was taken from eyeball to separate serum. Right after slaughtering, the liver and muscle were sampled to determine the deposit of protein, fat, cholesterol and biochemical parameters in serum. Result: (1) Every weekend, average weight of all GABA groups was higher than that of group CK (p＞0.05) and group C was highest at every stage, though the difference was not significant from other groups (p＞0.05). (2) Compared with group CK, the protein content of liver and muscle of each GABA group mice increased and group B was the highest, though the deposit difference in muscle was not significant among groups (p＞0.05). The deposit of protein in liver of group B was significantly higher than group CK (p＜0.01), A (p＜0.05) and C (p＜0.05). (3) Compared with group CK, the cholesterol content of liver and muscle of each GABA group decreased (p＞0.05) though without significant difference among GABA groups. The cholesterol content of group B was the lowest in liver while group C was the lowest in muscle. The content of fat in liver of each GABA group decreased (p＞0.05) though without significant difference among groups, while group B was the lowest and 2.25 percentage point lower than group CK. The deposit of fat in muscle enhanced, while group B was the highest, and that of group A (p＜0.05) and C (p＜0.05) were also higher than group CK. (4) Compared to group CK, the content of TP、ALB、GLOB、UA、TG、CHOL、HDL-C、LDL-C and activity of AKP、CHE、LDH、ALT in serum of GABA groups increased. The TP content of B was highest, and significantly different from group CK (p＜0.01). Group A and C were also significantly different from group CK (p＜0.05). The difference among GABA groups was not significant (p＞0.05). The ALB content of B was highest, and the difference among GABA groups was not significant (p＞0.05), though significantly higher than that of group CK (p＜0.05). The content of GLOB、UA、TG、HDL-C and activity of CK、CHE、LDH、ALT had no significant difference among different groups (p＞0.05). The GLOB、HDL-C content and CK activity of group B were highest. The content of UA and activity of CHE, LDH、ALT of group A were highest. The TG content of group C was highest. Compared to group CK, A/G value and GLU content of each GABA group reduced, but without significant difference among groups (p＞0.05). GLU and A/G were lowest for group A and group B, respectively. The content variation of BUN, CREA and BUN/CR was notegular while group C were higher than those of CK group (p＜0.05; p＞0.05; p＞0.05). The BUN content of group C was higher than that of group A (p＜0.05) and group B (p＜0.05), but the BUN content of group A and group B was lower than that of group CK (p＞0.05; p＞0.05; p＞0.05). CHOL content of group C (p＜0.01), A (p＜0.05) and B (p＜0.05) was higher than that of group CK, though the difference among GABA groups was not significant (p＞0.05). Compared to group CK, LDL-C content of group C (p＜0.01), B (p＜0.05), A (p＞0.05) and AKP of group C (p＞0.05), B (p＜0.05) and A (p＞0.05)all increased. Conclusion: GABA can enhance mice growth, strengthen the synthesis and deposit of protein in muscle and liver, reduce cholesterol deposition and content of fat in liver and also boost the deposition of fat in muscle.
发芽糙米是糙米发芽至适当长的芽体，主要由幼芽和带种皮的胚乳构成，富含许多生物活性物质如γ-氨基丁酸(gamma aminobutyric acid，GABA)，是一种具有全营养概念、特殊营养价值的功能性主食。本研究在糙米发芽后组织化学、理化特性的变化以及γ-氨基丁酸(GABA)富集探索基础上，进一步探讨GABA对小鼠体内糖类、脂类和蛋白质代谢的影响。旨在从宏观和微观揭示糙米发芽过程中发生的变化，评价发芽糙米的营养价值；筛选富含GABA发芽糙米制备的工艺条件；开发富含GABA发芽糙米及其制品。
1糙米发芽过程中营养成分及抗营养因子、碳水化合物组成及其相关酶活性的变化
目的：研究不同品种糙米发芽过程中营养成分和植酸含量、碳水化合物组成及相关酶活性的变化。方法：Ⅱ优838、川香优2号、江镇粳糯和香粳99糙米分别在35℃下发芽60h，每间隔12h取样并进行化学分析。结果：不同品种糙米发芽过程中淀粉、直链淀粉、支链淀粉和抗营养因子植酸含量均呈下降趋势，其他营养成分先上升后下降。总淀粉酶活力、α-淀粉酶活力先增加后降低。4个品种糙米发芽24h时抗坏血酸含量最高；赖氨酸含量约是发芽前的3倍，除江镇粳糯在发芽24h最高外，其余3个品种均在发芽36h达到峰值。色氨酸含量除Ⅱ优838在发芽48h最高外，其余3个品种在发芽36h达到峰值，约是发芽前的2倍。Ⅱ优838和川香优2号糙米可溶性蛋白、游离氨基酸和还原糖的含量在发芽36h达到峰值，江镇粳糯和香粳99的峰值则出现在发芽48h。其中，籼稻糙米(Ⅱ优838和川香优2号)糙米发芽36h时，还原糖和可溶性糖含量、总淀粉酶和α-淀粉酶活力达到高峰，而粳稻糙米(香粳99)和糯稻糙米(江镇粳糯)高峰值则出现在发芽48h时。结论：发芽可提高糙米营养成分的含量，降低抗营养因子植酸的含量；并通过影响糙米淀粉酶的活性，改善碳水化合物的组成，有利于糙米品质的改善。
2发芽对糙米蛋白质及氨基酸组成特性的影响
目的：研究发芽过程中糙米蛋白质和氨基酸含量和组成特性的变化，并与精米和糙米比较，评价发芽糙米蛋白质的营养价值。方法：糙米在35℃下发芽48h，每隔6h取样并进行化学分析。结果：发芽可以增加糙米蛋白质、总氨基酸和必需氨基酸的含量，必需氨基酸指数提高。发芽过程中清蛋白和谷蛋白含量均呈上升趋势，球蛋白含量呈下降的趋势。除发芽24h外，醇溶蛋白的含量在整个发芽期间也呈现增加趋势，增加的蛋白质主要是谷蛋白。各发芽阶段糙米的必需氨基酸和总氨基酸的比值均大于55％。18种氨基酸中有12种的含量在发芽24h最高。发芽后赖氨酸和苏氨酸的含量增加较明显，但仍是限制性氨基酸；丝氨酸、谷氨酸、精氨酸和组氨酸含量均有所增加。发芽24h时必需氨基酸指数最大，与FAO／WHO模式相比，必需氨基酸组成模式更加合理。结论：发芽可以提高糙米蛋白质的营养价值，改善籼稻米的品质，其中以发芽24h为宜。
3发芽对糙米淀粉理化特性和超微结构的影响
目的：研究发芽对糙米淀粉理化特性和超微结构的影响。方法：以未发芽、发芽12h、24h、36h的糙米为原料提取淀粉，并对淀粉及其组分含量、淀粉糊的透明度、凝沉性质、冻融稳定性、酶解率以及粘度等进行分析。结果：糙米发芽后淀粉及其组分的含量、淀粉糊的酶解率以及淀粉糊的粘度均降低；淀粉糊的透明度和冻融稳定性均增强；凝沉稳定性、糊化温度和淀粉颗粒超微结构变化不明显。结论：发芽对糙米淀粉的理化特性具有一定的改善作用。其中发芽24h的糙米淀粉的理化特性变化最明显。
4糙米发芽过程中GABA富集的研究
目的：研究不同浸泡剂对糙米发芽过程中γ-氨基丁酸富集的影响。方法：采用蒸馏水、CaCl_2溶液和壳聚糖溶液浸泡糙米，并通过控制不同的发芽温度(A：25℃、35℃、45℃)、发芽时间(B：12h、24h、36h)和pH(C：5.6、7.0、8.4)的正交试验设计[L_9(3~4)]，研究γ-氨基丁酸富集的适宜条件。结果：(1)蒸馏水浸泡的糙米在发芽24h前其GABA的含量呈现增加的趋势，发芽24h达到高峰，然后GABA含量呈下降趋势；各发芽阶段糙米中GABA含量均高于糙米和精米。(2)随着发芽时间的延长，0.1％CaCl_2溶液浸泡的糙米中GABA含量呈逐渐增加趋势，而0.5％CaCl_2溶液浸泡的糙米中GABA含量先增加后降低转而又增加，两种浓度下发芽42h时GABA均达到峰值，但低浓度下GABA的峰值高于高浓度。(3)随着发芽时间的延长，0.1％、0.3％和0.5％壳聚糖浸泡后糙米发芽24h内GABA含量呈逐渐增加趋势，24h后GABA含量逐渐减少；3种浓度下糙米发芽24h时GABA均达到峰值，在整个发芽期间，壳聚糖浓度高低与糙米中GABA含量呈正相关。(4)三因素三水平正交设计试验结果表明，pH为5.6时GABA富集效果明显。各因素水平对GABA含量的影响从主到次的顺序为C＞A＞B。结合前面系列实验结果综合考虑，选择各因素的最佳水平组合为A_3B_2C_1，即发芽温度为45℃、发芽时间为24h、发芽pH为5.6。
结论：蒸馏水、低浓度的CaCl_2和壳聚糖浸泡均有利于糙米发芽过程GABA的富集；pH值是影响发芽糙米GABA富集的主要因素，其中酸性环境有利于GABA的富集，其次是发芽温度，再次是发芽时间。
5小鼠体内GABA生理功效的研究
目的：通过分析GABA对小鼠体内血清、肌肉和肝脏中与糖类、脂类和蛋白质等代谢相关的生理生化指标的影响，探讨小鼠体内GABA的生理功效。方法：56只体重22g左右的4w雄性昆明种小鼠(p＞0.05)，随机分成4组，每组设2个重复，每个重复7只，分别为CK组(对照组，GABA量为零)、A组(低剂量组，GABA量为12.5mg／L)、B组(中剂量组，GABA量为125mg／L)和C组(高剂量组，GABA量为1250mg／L)。饲养2d后每天早晨0.2mL／只空腹灌胃，连续18d；每周末清晨空腹称重一次；屠宰前禁食12h，眼球取血，分离血清；并迅速处死，取其肝脏和肌肉，分析肝脏和肌肉中蛋白质、脂肪和胆固醇的沉积以及血清生理生化指标。结果：(1)每周末各GABA组小鼠的平均体重均高于CK组(p＞0.05)；C组小鼠的平均体重最大，但与其他各组间差异不显著(p＞0.05)。(2)与CK组相比，各GABA组小鼠肝脏和肌肉中蛋白质含量均增加，以B组肝脏和肌肉中蛋白质含量最高，但各组小鼠肌肉中蛋白质的沉积差异不显著(p＞0.05)；而B组肝脏中蛋白质的沉积显著高于CK组(p＜0.01)、A组(p＜0.05)和C组(p＜0.05)。(3)与CK组相比，各GABA组小鼠肝脏和肌肉中胆固醇含量均下降，但各组间差异不显著(p＞0.05)。其中肝脏胆固醇以B组最低，而肌肉中以C组最低。各GABA组小鼠肝脏脂肪含量也均有所降低，但各组间差异不显著(p＞0.05)，其中以B组最低，比对照组降低2.25个百分点；而肌肉中脂肪的沉积却加强，其中B组沉积量最大，高于CK组(p＜0.01)、A组(p＜0.05)和C组(p＜0.05)。(4)与CK组相比，各GABA组小鼠血清中TP、ALB、GLOB、UA、TG、CHOL、HDL-C、LDL-C含量以及AKP、CHE、LDH、ALT的活性均增加。其中B组TP最高，与CK组间差异极显著(p＜0.01)，A组和C组与CK组间差异显著(p＜0.05)，各GABA组间差异不显著性(p＞0.05)。ALB以B组最高，各GABA组间无显著性差异(p＞0.05)，但均显著高于CK组(p＜0.05)。各组的GLOB、UA、TG、HDL-C的含量以及CK、CHE、LDH、ALT的活性间差异不显著(p＞0.05)，其中B组的GLOB、HDL-C和CK最高，A组的UA、CHE、LDH、ALT最高，TG以C组最高。各GABA组小鼠的A／G、GLU均比CK组有所下降，但各组间差异不显著(p＞0.05)，其中A组的GLU最低，B组的A／G最低。各GABA组的BUN、CREA、BUN／CR与CK组相比增减不一，其中C组的这三个指标均高于CK组(p＜0.05；p＞0.05；p＞0.05)，而A组和B组的均低于CK组(p＞0.05；p＞0.05；p＞0.05)，此外，C组的BUN也高于A组(p＜0.05)和B组(p＜0.05)。C组(p＜0.01)、A组(p＜0.05)和B组(p＜0.05)的CHOL均高于CK组，但A、B、C组间无显著性差异(p＞0.05)。与CK组相比，C组(p＜0.01)、B组(p＜0.05)和A组(p＞0.05)的LDL-C均增加，A组(p＞0.05)、C组(p＞0.05)和B组(p＜0.05)的AKP也增加。结论：GABA可促进小鼠生长；加强肌肉和肝脏中蛋白质的合成与沉积，减少胆固醇的沉积，降低肝脂含量，加强肌脂的沉积。
Brown rice dehulled from paddy has poor taste, lower cooking capability and high fat content though enriched with nutrition that, however, tends to decay because of acidification for long term stocking. Substantial nutrition loss will happen during the process from brown rice to polished one. Thus it will be the trend that conventional rice processing techniques combine with modern biological farther processing methods.
Germinated brown rice consists of brown rice with proper amount of sprout and endosperm with seed coat, which is enriched with many biological active components such as gamma aminobutyric acid (GABA) and thus regarded as functional food that combines broad spectrum nutrition with special nutrient. With the study of histochemical change, physicochemical property, and GABA accumulation during germinating of brown rice, the effect is farther investigated of GABA on the metabolism of carbohydrate, lipid and protein of mice. This investigation was aimed to reveal microcosmic and macroscopic variation of brown rice during germination, evaluate its nutrition, determine the optimum germinating condition and develop germinated brown rice products enriched with GABA.
1 Investigation of brown rice germination on nutrients, anti-nutrition factor, carbohydrate and related enzyme
Purpose: Investigation of germination on nutrients, phytic acid, carbohydrate content and related enzyme activity in brown rice. Method: Four types of brown rice (Ⅱyou838, Chuanxiangyou2, Xiangjing99, Jiangzhenjingnuo) were selected to germinate for 60 hours under 35℃. Samples were taken for each 1:2 hours and tested chemically. Result: For all types, tests revealed that the contents of starch, amylose, amylopectin and anti-nutrition factor phytic acid reduced, while other nutrition content, activity of total amylase andα-amylase increased at first and then reduced. For all four types, ascorbic acid contents reached the highest level at 24h, and the lysine contents were about 3 times higher than those before germination and reached the highest level at 36h except for Jiangzhenjingnuo whose apex was at 24 hours. Tryptophan content reached the highest level at 36h-about 2 times of original except forⅡyou838 whose apex was at 48h. The contents of soluble protein, free amino acid and reduced sugar inⅡyou838 and Chuanxiangyou2 reached the highest level at germination of 36h, while the highest values happened at germination of 48h for Xiangjing99 and Jiangzhenjingnuo. For indica type rice among them (Ⅱyou838 and Chuanxiangyou2), the max. contents of reduced and soluble sugar, total amylase and a-amylase activity happened at germinated 36h, while 48h for nonglutinous rice (Xiangjing99) and glutinous rice (Jiangzhenjingnuo). Conclusion: By germinating, the contents of nutrition increased and phytic acid as anti-nutrition factor reduced. Besides, carbohydrate composition and the quality of brown rice were improved through affecting the amylase activity.
2 Effect of germination on protein and amino acid composition properties of brown rice
Purpose: Assessing the protein nutrition value of germinated brown rice by investigating the content and composition variation of protein and amino acid during germination, comparing with brown rice and polished rice as reference. Method: Brown rice was kept at 35℃for 48 hours and samples were taken every 6h and tested chemically. Result: Germination can increase the content of crude protein (CP), total amino acid (TAA), essential amino acid (EAA) and its index (EAAI). Content of albumin and glutelin increased as well while dropping of globulin content during germination. Except germinating for 24 hours, the content of prolamine rose and mostly was in glutelin. During the process, the ratio of essential amino acid to total amino acid kept over 55%. 12 of 18 types of amino acid reached the highest level at germination of 24 hours. The content of lysine and the threonine increased substantially also after germination though as limited amino acid yet. The content of serine, glutamic acid, arginine and histidine all increased then. EAAI summited at germination of 24 hours and with had more preferable EAA composition compared with FAO/WHO. Conclusion: The nutritional value of protein and the quality of indica brown rice can be improved by germinating and 24h is the most preferred germinating time.
3 Effect of germination on starch physicochemieal properties and granular micro-structure in brown rice
Purpose: Investigation of the effect of germination on starch physicochemical property and granular micro-structure of brown rice. Method: Extracting starch from brown rice germinated for Oh, 12h, 24h and 36h and testing its starch content, components, paste clarity, retrogradation, freeze-thaw stability, enzymatic hydrolyzation and viscosity. Result: After germinating of brown rice, the content of starch and its component, enzymatic hydrolyzation and viscosity declined, whereas clarity and freeze-thaw stability enhanced. Paste retrogradation, gelatinization temperature and granular micro-structure of starch had no obvious change. Conclusion: Germinating resulted in improvement of physicochemical properties of brown rice starch and that at 24h was most significant.
4 Accumulation in brown rice of GABA during the germination
Purpose: Investigation of the effect of different soaking agents on the accumulation of gamma aminobutyric acid in germinated brown rice. Method: Soaking brown rice with distilled water, CaCl_2 solution and chitosan at different temperature (A: 25℃, 35℃, 45℃), time (B: 12h, 24h, 36h) and pH (C: 5.6, 7.0, 8.4) that was designed as three-factor and three-level orthogonal experiment [L_9 (3_4)] to determined the content variation of GABA in the process of brown rice germination. Result: (1) Within 24h, the GABA content in brown rice soaked in distilled water increased, then decreased and kept higher than that of brown rice and polish rice in every germination stage. (2) Along with the germination time, the GABA content of brown rice soaked in 0.1%CaCl_2 solution gradually increased, while that in 0.5%CaCl_2 solution increased at first and then reduced and increased again. The peak of GABA all happened at germination of 42h, and was higher in low concentration than that in high one. (3) Along with the germination time, the GABA content of brown rice soaked in 0.1%、0.3%、0.5% chitosan solution gradually increased within 24h then decreased. The peaks of GABA all happened at germination 24h. The content of GABA was positively correlated with that of chitosan. (4) The orthogonal experiment revealed that GABA accumulated substantially at pH5.6. Factors affecting GABA content ranked as C＞A＞B. Considering results of all series as a whole, the optimum condition of GABA accumulation in the process of brown rice germination was A_3B_2C_1, namely 45℃, 24h and pH5.6. Conclusion: Distilled water, low concentration CaCl_2 solution and chitosan all can enhance GABA accumulation of brown rice in germination, pH is the key factor affecting GABA accumulation, while second is germination temperature and time the third. Acid condition can promote GABA accumulation.
5 Physiological functions of GABA on mice
Purpose: Investigation of physiological functions of GABA on mice by analyzing the effects on biochemical parameters correlative to the metabolism of carbohydrate, lipid and protein in mice serum, muscle and liver. Method: 56 male Kunming mice of 4-w-old and weighing about 22g (p＞0.05) were randomly divided into 4 groups, and each group consisted of 14, and 7 of per group serving for one repeat. The groups were named as group CK (Control group, 0mg/L GABA), group A (low dose, 12.5mg/L GABA), group B (middle dose, 125mg/L GABA) and group C (high dose, 1250mg/L GABA). After two day's breeding, each mouse was filled in stomach with 0.2mL GABA for 18d at limosis state every morning, and was weighed every weekend morning under limosis state. With 12-hour fasting before slaughter, the blood sample was taken from eyeball to separate serum. Right after slaughtering, the liver and muscle were sampled to determine the deposit of protein, fat, cholesterol and biochemical parameters in serum. Result: (1) Every weekend, average weight of all GABA groups was higher than that of group CK (p＞0.05) and group C was highest at every stage, though the difference was not significant from other groups (p＞0.05). (2) Compared with group CK, the protein content of liver and muscle of each GABA group mice increased and group B was the highest, though the deposit difference in muscle was not significant among groups (p＞0.05). The deposit of protein in liver of group B was significantly higher than group CK (p＜0.01), A (p＜0.05) and C (p＜0.05). (3) Compared with group CK, the cholesterol content of liver and muscle of each GABA group decreased (p＞0.05) though without significant difference among GABA groups. The cholesterol content of group B was the lowest in liver while group C was the lowest in muscle. The content of fat in liver of each GABA group decreased (p＞0.05) though without significant difference among groups, while group B was the lowest and 2.25 percentage point lower than group CK. The deposit of fat in muscle enhanced, while group B was the highest, and that of group A (p＜0.05) and C (p＜0.05) were also higher than group CK. (4) Compared to group CK, the content of TP、ALB、GLOB、UA、TG、CHOL、HDL-C、LDL-C and activity of AKP、CHE、LDH、ALT in serum of GABA groups increased. The TP content of B was highest, and significantly different from group CK (p＜0.01). Group A and C were also significantly different from group CK (p＜0.05). The difference among GABA groups was not significant (p＞0.05). The ALB content of B was highest, and the difference among GABA groups was not significant (p＞0.05), though significantly higher than that of group CK (p＜0.05). The content of GLOB、UA、TG、HDL-C and activity of CK、CHE、LDH、ALT had no significant difference among different groups (p＞0.05). The GLOB、HDL-C content and CK activity of group B were highest. The content of UA and activity of CHE, LDH、ALT of group A were highest. The TG content of group C was highest. Compared to group CK, A/G value and GLU content of each GABA group reduced, but without significant difference among groups (p＞0.05). GLU and A/G were lowest for group A and group B, respectively. The content variation of BUN, CREA and BUN/CR was notegular while group C were higher than those of CK group (p＜0.05; p＞0.05; p＞0.05). The BUN content of group C was higher than that of group A (p＜0.05) and group B (p＜0.05), but the BUN content of group A and group B was lower than that of group CK (p＞0.05; p＞0.05; p＞0.05). CHOL content of group C (p＜0.01), A (p＜0.05) and B (p＜0.05) was higher than that of group CK, though the difference among GABA groups was not significant (p＞0.05). Compared to group CK, LDL-C content of group C (p＜0.01), B (p＜0.05), A (p＞0.05) and AKP of group C (p＞0.05), B (p＜0.05) and A (p＞0.05)all increased. Conclusion: GABA can enhance mice growth, strengthen the synthesis and deposit of protein in muscle and liver, reduce cholesterol deposition and content of fat in liver and also boost the deposition of fat in muscle.
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