醋酸菌优良菌株的筛选及培养基优化
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摘要
醋酸菌是醋酸发酵的主要工业用菌。国外非常重视对醋酸菌优势菌株的选育和醋酸发酵机理的研究,也做了大量工作。国内对醋酸菌优良菌种的应用也日益重视起来,并积累了许多有关醋酸菌分类鉴定及性能研究的资料。目前国内应用纯培养醋酸菌的食醋酿造厂家多采用AS1.41号进行醋酸发酵,在稳产方面取得了较好的效果,但是将其应用于果醋生产方面,不仅产酸能力、耐酒精能力有待提高,而且形成的风味也不佳。因此,寻找有利于果醋生产的特定菌株成为当前研究的重点。为了拥有自主知识产权的醋酸菌株,本研究首先从实验室自然发酵的果醋醪和果醋液中分离筛选出了一株性能优良的产酸菌株F_9,对菌株自身特性进行研究后,优化了发酵工艺参数,为工业化生产奠定了基础;同时对产酸促进因子进行了研究,为醋酸菌株的进一步研究和应用提供了理论指导。本文得到的主要结论如下:
①按常规菌种分离筛选的方法,以A_S1.41为对照菌株,从自然发酵的果醋醪和果醋液中分离出了高活性、高产酸能力的醋酸菌菌株F_9,经过对菌株的形态和生理生化特征的分析,初步鉴定菌株F_9为醋化醋杆菌菌种。
②确立了适合菌株F_9生长繁殖的种子培养基和培养条件。种子培养基最优组成为:葡萄糖2%,酵母膏2%,无水乙醇3%,KH2PO_40.05%,MgSO_40.3%;最佳种子培养条件为:种龄36h、温度30℃、摇床转速150r/min。
③确立了适合菌株F_9液态发酵产酸的发酵培养基。发酵培养基的配方为:葡萄糖1%,酵母膏1.5%,KH_2PO_40.05%,MgSO_40.3%,无水乙醇5.63%,以此优化培养基获得总酸的产量是基础发酵培养基的1.20倍。
④将几种因子添加到发酵培养基中,研究其对菌株F_9发酵产酸的影响,并对其最适添加量进行了试验。结果表明抗坏血酸、葡萄糖酸、乳酸、乙酸四种因子对菌株F_9发酵产酸均有一定的促进作用,当抗坏血酸1%时产酸增幅可达到1.33倍,葡萄糖酸的添加量为2.5%时,产酸增幅可达到1.44倍。
The acetic acid bacterium is a major bacteria in fermentation industry. Many researchers aboard not only pay more attention to screen superior strains of acetic acid bacteria but also have done a lot of research works on the fermentation mechanism of acetic acid. In our country the application of acetic acid bacteria is also paid attention gradually, and a lot of information on isolation、identification and properties of acetic acid bacteria is accumulated. At present, vinegar brewing manufacturers who applied pure culture of acetic acid bacteria to acetic acid fermentation mostly use AS1.41, which has achieved the stable production, but if used in the production of the fruit vinegar, it is the improvement required of the ability acid-poduction and alcohol-resistant and the formation of poor taste. Therefore, looking for a particular strain favoring fruit vinegar production has become the focus of current research.
In order to have acetic acid strains of proprietary intellectual property rights, the strain (F_9) with high yield of acetic acid was initially selected from fruit vinegar and then the characteristics of the strain F_9 was researched, the parameters of fermentation process were optimized which established the basis for industry. Acid production-enhancing factors were observed which provided theoretical guidance for the further research on acetic acid strains. The results of this research were as follows:
①A strain (F_9) with the ability of high-activity and high acid production was selected from the fruit mash and vinegar solution of natural fermentation by conventional methods, with AS1.41 for the contrast strain. After the configuration and physiological and biochemical characteristics were studied, the strain was primarily ascertained to belong to Acetobacter acetic.
②The seed medium and seed cultivation conditions were confirmed. Optimal composition of the seed medium was as follows: glucose 2%, yeast extract 2%, ethanol 3%, KH_2PO_4 0.05%, MgSO_4 0.3%. The optimum cultivation conditions were seed age 36h, temperature 30℃, rotation speed 150r/min.
③The liquid fermentation medium was confirmed. The components of it were as follows: glucose 1%, yeast extract 1.5%, KH_2PO_4 0.05%, MgSO_40.3%, ethanol 5.63%. Under this fermentation medium, the yield of acetic acid was as 1.20 times as that under initial fermentation medium.
④Several factors has been added to the fermentation medium to study the impact of acid fermentation on the strain F_9, and its optimal dosage was tested. Four factors, namely ascorbic acid, gluconic acid, lactic acid and acetic acid were determined which had promotation for fermentation. Ascorbic acid was added 1%, the growth rate could reach 1.33 times , gluconic acid was added 2.5%, the growth rate could reach 1.44 times.
①按常规菌种分离筛选的方法,以A_S1.41为对照菌株,从自然发酵的果醋醪和果醋液中分离出了高活性、高产酸能力的醋酸菌菌株F_9,经过对菌株的形态和生理生化特征的分析,初步鉴定菌株F_9为醋化醋杆菌菌种。
②确立了适合菌株F_9生长繁殖的种子培养基和培养条件。种子培养基最优组成为:葡萄糖2%,酵母膏2%,无水乙醇3%,KH2PO_40.05%,MgSO_40.3%;最佳种子培养条件为:种龄36h、温度30℃、摇床转速150r/min。
③确立了适合菌株F_9液态发酵产酸的发酵培养基。发酵培养基的配方为:葡萄糖1%,酵母膏1.5%,KH_2PO_40.05%,MgSO_40.3%,无水乙醇5.63%,以此优化培养基获得总酸的产量是基础发酵培养基的1.20倍。
④将几种因子添加到发酵培养基中,研究其对菌株F_9发酵产酸的影响,并对其最适添加量进行了试验。结果表明抗坏血酸、葡萄糖酸、乳酸、乙酸四种因子对菌株F_9发酵产酸均有一定的促进作用,当抗坏血酸1%时产酸增幅可达到1.33倍,葡萄糖酸的添加量为2.5%时,产酸增幅可达到1.44倍。
The acetic acid bacterium is a major bacteria in fermentation industry. Many researchers aboard not only pay more attention to screen superior strains of acetic acid bacteria but also have done a lot of research works on the fermentation mechanism of acetic acid. In our country the application of acetic acid bacteria is also paid attention gradually, and a lot of information on isolation、identification and properties of acetic acid bacteria is accumulated. At present, vinegar brewing manufacturers who applied pure culture of acetic acid bacteria to acetic acid fermentation mostly use AS1.41, which has achieved the stable production, but if used in the production of the fruit vinegar, it is the improvement required of the ability acid-poduction and alcohol-resistant and the formation of poor taste. Therefore, looking for a particular strain favoring fruit vinegar production has become the focus of current research.
In order to have acetic acid strains of proprietary intellectual property rights, the strain (F_9) with high yield of acetic acid was initially selected from fruit vinegar and then the characteristics of the strain F_9 was researched, the parameters of fermentation process were optimized which established the basis for industry. Acid production-enhancing factors were observed which provided theoretical guidance for the further research on acetic acid strains. The results of this research were as follows:
①A strain (F_9) with the ability of high-activity and high acid production was selected from the fruit mash and vinegar solution of natural fermentation by conventional methods, with AS1.41 for the contrast strain. After the configuration and physiological and biochemical characteristics were studied, the strain was primarily ascertained to belong to Acetobacter acetic.
②The seed medium and seed cultivation conditions were confirmed. Optimal composition of the seed medium was as follows: glucose 2%, yeast extract 2%, ethanol 3%, KH_2PO_4 0.05%, MgSO_4 0.3%. The optimum cultivation conditions were seed age 36h, temperature 30℃, rotation speed 150r/min.
③The liquid fermentation medium was confirmed. The components of it were as follows: glucose 1%, yeast extract 1.5%, KH_2PO_4 0.05%, MgSO_40.3%, ethanol 5.63%. Under this fermentation medium, the yield of acetic acid was as 1.20 times as that under initial fermentation medium.
④Several factors has been added to the fermentation medium to study the impact of acid fermentation on the strain F_9, and its optimal dosage was tested. Four factors, namely ascorbic acid, gluconic acid, lactic acid and acetic acid were determined which had promotation for fermentation. Ascorbic acid was added 1%, the growth rate could reach 1.33 times , gluconic acid was added 2.5%, the growth rate could reach 1.44 times.
引文
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