Effects of solid-state fermentation of Hongqu rice wine grains on spore production and enzyme-production characteristics of Aspergillus oryzae 3.042
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摘要:
目的 研究米曲霉3.042红曲糟固态制曲产孢子及产酶特性，为米曲霉固态发酵红曲糟制备调味液提供理论支持。 方法 以红曲糟为主要原料，以米曲霉3.042为发酵剂，以酸性蛋白酶、中性蛋白酶、碱性蛋白酶酶活力及蛋白质酶解率、氨氮转化率等为指标，研究红曲糟、麸皮与水做培养基料配比、发酵时间等因素对米曲霉生长及产酶效果的影响。 结果 成曲培养料适宜质量配比为红曲糟∶麸皮∶水为1∶0.8∶1.4，在此条件下培养72 h，成曲的孢子量、酸性蛋白酶、碱性蛋白酶、中性蛋白酶和谷氨酰胺酶分别可达9.93、1160.94 U·g−1、1157.47 U·g−1、1788.40 U·g−1和10.49 U·g−1。利用该成曲发酵得到的调味液中氨基酸态氮、酶解率和氨氮转化率分别为 8.2 g·L−1、63.40%和37.08%，利用红曲糟麸皮做培养基料，利用3.042发酵得到的调味液鲜味较高，色泽红褐有光泽，具备酱香浓郁、滋味醇厚的特征，具有典型的酱油风味。 结论 红曲糟适合做米曲霉固态制曲的培养基料的主要原料，其适宜的质量比为红曲糟∶麸皮∶水为1∶0.8∶1.4。 Abstract:Objective In this paper, the characteristics of spore production and enzyme production of Aspergillus oryzae 3.042 from solid-state koji-making Hongqu rice wine grains were studied in order to provide theoretical support for the preparation of seasoning liquid from Aspergillus oryzae solid-state fermentation of Hongqu rice wine grains. Method Hongqu rice wine grains was used as the main raw material. Aspergillus oryzae strain 3.042 was used as the starter. The enzyme activities of acidic, neutral and alkaline proteases, protein enzymatic hydrolysis rate, and ammonia nitrogen conversion rate were used as indicators. The researches on Hongqu rice wine grains and bran were studied to make the effects of culture material ratio, fermentation time and other factors on the growth and enzyme production of Aspergillus oryzae clear. Results The suitable ratio of koji culture material is Hongqu rice wine grains∶bran∶water = 1∶0.8∶1.4. Cultured under this condition for 72 h, the amount of spores, acid protease, alkaline protease, neutral protease of koji , glutaminase were up to 9.93, 1160.94 U·g−1, 1157.47 U·g−1, 1788.40 U·g−1, 10.49 U·g−1. The amino acid nitrogen, enzymatic hydrolysis rate and ammonia nitrogen conversion rate in the crude oil obtained by the koji fermentation were 8.2 g·L−1, 63.40% and 37.08% respectively.Used Hongqu rice wine grains as culture material ratio. The crude sauce has a high flavor, red brown color and luster. The sauce is rich in flavor and has a typical soy sauce style. Conclusion Hongqu rice wine grains are suitable for solid-state koji-making by Aspergillus oryzae, and the suitable ratio is bran:Hongqu rice wine grains:water =1:0.8:1.4. -
图 1 不同培养料配比对米曲霉3.042产孢子的影响
Figure 1. Effects of different culture material ratio on sporulation of Aspergillus oryzae 3.042
图 2 不同培养料配比对米曲霉3.042产蛋白酶的影响
酸性：酸性蛋白酶酶活力；碱性：碱性蛋白酶酶活力；中性：中性蛋白酶酶活力。
Figure 2. Effect of different culture material ratio on protease of Aspergillus oryzae 3.042
Acid: acid protease enzyme activity; alkaline: alkaline protease enzyme activity; neutral: neutral proteinase enzyme activity.
图 3 米曲霉3.042红曲糟固态制曲过程孢子量的变化
Figure 3. Number of spores changes in the Hongqu rice wine grains solid-state fermentation of Aspergillus oryzae 3.042
图 4 米曲霉3.042红曲糟固态制曲过程不同蛋白酶变化趋势
Figure 4. Trends of different protease in the Hongqu rice wine grains solid-state fermentation of Aspergillus oryzae 3.042
表 1 酱油原油的感官评分标准
Table 1. Sensory scoring criteria for crude sauce
色泽（25分）Tincture（25） 酱香味（25分）Sauce fragrance（25） 滋味（25分）taste（25） 香气（25分）sweet smell（25） 红褐色，颜色较深有光泽（16~25）Red-brown, darker and shiny（16~25） 具有酱油典型性风格（16~25）With a typical style of soy sauce（16~25） 滋味感强，容易入口（16~25）Strong sense of taste, easy to enter （16~25） 有明显香味（16~25）Obvious fragrance（16~25） 颜色过深，有少许沉淀（5~15）The color is too dark, with a little precipitation（5~15） 酱油味风格一般（5~15）Soy sauce flavor style is mediocre（5~15） 滋味感一般（5~15）Average taste（5~15） 香味一般，容易接受（5~15）General fragrance, easy to accept（5~15） 颜色浅，无光泽（＜5）Light in color, dull and dull（＜5） 没有酱油风味（＜5）No soy sauce flavor（＜5） 滋味感较差（＜5）Poor taste（＜5） 没有香味，刺激味强（＜5）No fragrance, stimulate the strong taste（＜5） 
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表 2 不同培养料配比对米曲霉3.042酶解发酵指标的影响
Table 2. Effect of different culture material ratio on fermentation indexof Aspergillus oryzae 3.042
处理组Processing group 酶解率The enzymatic hydrolysis rate/% 氨氮转化率Conversion rate of ammonia nitrogen/% 1∶0.6∶1.2 58±0.5 Ab 35.87±0.35 Ab 1∶0.8∶1.4 63.4±2.8 Aa 37.08±0.71 Aa 1∶1∶1.5 59.6±1.9 Aab 36.11±0.23 Aab 1∶1.2∶1.6 47.3±3.2 Bc 32.98±1.22 Bc 1∶1.4∶1.8 42.1±1.0 Bd 31.21±0.88 Bd 不同大写字母指组间差异呈极显著水平(P＜0.01)；不同小写字母指组间差异呈显著水平(P＜0.05)。下同。Different capital letters indicate extremely significant difference between treatments (P ＜0.01); different lower case letters indicate significant difference between treatments (P ＜0.05). Same for below.
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表 3 成曲的质量分析
Table 3. Quality analysis of finished koji
指标/target 孢子数对数值Spore count 酸性蛋白酶acid protease/（U·g−1） 碱性蛋白酶alkali protease/（U·g−1） 中性蛋白酶neutral protease/（U·g−1） 谷氨酰胺酶glutaminase/（U·g−1） 水分含量humidity/% pH值pH 成曲/finished koji 9.93±0.05 1160.94±25.30 1157.47±25.20 1516.84±15.84 10.49±0.33 26.04±0.55 6.5
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表 4 酱油原油的主要指标
Table 4. Main indicators of crude sauce
指标/target 酱油原油/crude sauce 酶解率 The enzymatic hydrolysis rate /% 63.4±2.8 氨氮转化率 Conversion rate of ammonia nitrogen/ % 37.08±0.71 氨基酸态氮amino acid nitrogen/（ g·L−1） 8.2±0.4
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