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        地衣芽孢桿菌WX—02補(bǔ)硝酸鈉發(fā)酵產(chǎn)聚γ—谷氨酸工藝

        2014-05-04 18:33:44許尚華胡麗芳陳守文
        湖北農(nóng)業(yè)科學(xué) 2014年4期
        關(guān)鍵詞:硝酸鈉華中農(nóng)業(yè)大學(xué)補(bǔ)料

        許尚華++胡麗芳++陳守文

        摘要:以地衣芽孢桿菌(Bacillus licheniformis)WX-02為研究對(duì)象,以提高其發(fā)酵產(chǎn)生的聚γ-谷氨酸的產(chǎn)量為研究目的,經(jīng)搖瓶發(fā)酵得到優(yōu)化的硝酸鈉補(bǔ)料方案,即初始硝酸鈉濃度為10 g/L,發(fā)酵至12 h時(shí)補(bǔ)加5 g/L硝酸鈉時(shí)聚γ-谷氨酸產(chǎn)量最大。將此補(bǔ)料方案在3 L發(fā)酵罐上進(jìn)行驗(yàn)證。結(jié)果表明,發(fā)酵得到的菌體生物量和聚γ-谷氨酸產(chǎn)量最大值分別為4.5 g/L和38.7 g/L,與對(duì)照相比分別提高55.2%和14.8%。通過補(bǔ)硝酸鈉可以提高菌體生物量和對(duì)谷氨酸的利用率,從而提高聚γ-谷氨酸的發(fā)酵水平。

        關(guān)鍵詞:地衣芽孢桿菌(Bacillus licheniformis);聚γ-谷氨酸;硝酸鈉;補(bǔ)料發(fā)酵;生物量

        中圖分類號(hào):TQ920.6 文獻(xiàn)標(biāo)識(shí)碼:A 文章編號(hào):0439-8114(2014)04-0903-04

        Optimizing Sodium Nitrate Fed-batch Culture on Poly(γ-glutamic acid) Fermentation by Bacillus licheniformis WX-02

        XU Shang-hua,HU Li-fang,CHEN Shou-wen

        (Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University , Wuhan 430070,China)

        Abstract: Bacillus licheniformis WX-02 was used to study the ways of improving poly(γ-glutamic acid)(γ-PGA) yield. The optimal conditions of sodium nitrate feeding for γ-PGA fermentation by Bacillus licheniformis WX-02 by flask-shaking fed-batch fermentation were the initial sodium nitrate concentration 10 g/L, and 5 g/L of sodium nitrate added when fermentation to 12 h. The fermentation was carried out in a 3 L fermentor, showing that the maximum of biomass and γ-PGA yield was 4.5 g/L and 38.7 g/L with increase of 55.2% and 14.8%, compared with the control. It is indicated that adding sodium nitrate can increase biomass, enhance the utilization of glutamate, and improve the γ-PGA fermentation.

        Key words: Bacillus licheniformis; poly-gamma-glutamic acid; sodium nitrate; feeding fermentation; biomass

        1 材料與方法

        1.1 材料

        1.1.1 菌株 地衣芽孢桿菌WX-02,由華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物學(xué)國家重點(diǎn)實(shí)驗(yàn)室篩選和保藏,保藏號(hào)為CCTCC M208065。

        1.2 方法

        參考文獻(xiàn):

        [1] 陳詠竹,孫啟玲.γ-多聚谷氨酸的性質(zhì)、發(fā)酵生產(chǎn)及其應(yīng)用[J].微生物學(xué)通報(bào),2004,31(1): 122-126.

        [2] BAJAJ I, SINGHAL R. Poly(glutamic acid)- an emerging biopolymer of commercial interest[J]. Bioresour Technol,2011, 102(10):5551-5561.

        [3] 曹名鋒,金映虹,解 慧,等.γ-聚谷氨酸的微生物合成、相關(guān)基因及應(yīng)用展望[J].微生物學(xué)通報(bào),2011,38(3):388-395.

        [4] 蔡 謹(jǐn),孫章輝,王 雋,等.補(bǔ)料發(fā)酵工藝的應(yīng)用及其研究進(jìn)展[J].工業(yè)微生物,2005,35(1):42-48.

        [5] 呂 萌,梁金鐘,王風(fēng)青.補(bǔ)料發(fā)酵枯草芽孢桿菌合成γ-聚谷氨酸的研究[J].食品科學(xué),2011, 32(23):225-228.

        [6] 胡麗芳,李 欣,冀志霞,等.地衣芽胞桿菌WX-02補(bǔ)糖發(fā)酵聚γ-谷氨酸的工藝優(yōu)化[J]. 華中農(nóng)業(yè)大學(xué)學(xué)報(bào),2012,31(3):287-292.

        [7] YOON S H, DO J H, LEE S Y, et al. Production of poly-γ-glutamic acid by fed-batch culture of Bacillus licheniformis[J]. Biotechnology Letters,2000,22(7):585-588.

        [8] KRAFTA B, STROUSA M, TEGETMEYER H E. Microbial nitrate respiration -Genes, enzymes and environmental distribution[J]. Journal of Biotechnology, 2011,155(1):104-117.

        [9] CHENG C, ASADA Y, AIDA T. Production of γ-polyglutamic acid by Bacillus licheniformis A35 under denitrifying conditions[J]. Agricultural and Biological Chemistry,1989,53(9):2369-2375.

        [10] XU K D, STEWART P S, XIA F, et al. Spatial physiological heterogeneity in Pseudomonas aeruginosa biofilm is determined by oxygen availability[J]. Appl Environ Microbiol, 1998,64(10):4035-4039.

        [11] WEI X T, JI Z X, CHEN S W. Isolation of halotolerant Bacillus licheniformis WX-02 and regulatory effects of sodium chloride on yield and molecular sizes of poly-γ-glutamic acid[J]. Appl Biochem Biotechnol,2010,160(5):1332-1340.

        摘要:以地衣芽孢桿菌(Bacillus licheniformis)WX-02為研究對(duì)象,以提高其發(fā)酵產(chǎn)生的聚γ-谷氨酸的產(chǎn)量為研究目的,經(jīng)搖瓶發(fā)酵得到優(yōu)化的硝酸鈉補(bǔ)料方案,即初始硝酸鈉濃度為10 g/L,發(fā)酵至12 h時(shí)補(bǔ)加5 g/L硝酸鈉時(shí)聚γ-谷氨酸產(chǎn)量最大。將此補(bǔ)料方案在3 L發(fā)酵罐上進(jìn)行驗(yàn)證。結(jié)果表明,發(fā)酵得到的菌體生物量和聚γ-谷氨酸產(chǎn)量最大值分別為4.5 g/L和38.7 g/L,與對(duì)照相比分別提高55.2%和14.8%。通過補(bǔ)硝酸鈉可以提高菌體生物量和對(duì)谷氨酸的利用率,從而提高聚γ-谷氨酸的發(fā)酵水平。

        關(guān)鍵詞:地衣芽孢桿菌(Bacillus licheniformis);聚γ-谷氨酸;硝酸鈉;補(bǔ)料發(fā)酵;生物量

        中圖分類號(hào):TQ920.6 文獻(xiàn)標(biāo)識(shí)碼:A 文章編號(hào):0439-8114(2014)04-0903-04

        Optimizing Sodium Nitrate Fed-batch Culture on Poly(γ-glutamic acid) Fermentation by Bacillus licheniformis WX-02

        XU Shang-hua,HU Li-fang,CHEN Shou-wen

        (Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University , Wuhan 430070,China)

        Abstract: Bacillus licheniformis WX-02 was used to study the ways of improving poly(γ-glutamic acid)(γ-PGA) yield. The optimal conditions of sodium nitrate feeding for γ-PGA fermentation by Bacillus licheniformis WX-02 by flask-shaking fed-batch fermentation were the initial sodium nitrate concentration 10 g/L, and 5 g/L of sodium nitrate added when fermentation to 12 h. The fermentation was carried out in a 3 L fermentor, showing that the maximum of biomass and γ-PGA yield was 4.5 g/L and 38.7 g/L with increase of 55.2% and 14.8%, compared with the control. It is indicated that adding sodium nitrate can increase biomass, enhance the utilization of glutamate, and improve the γ-PGA fermentation.

        Key words: Bacillus licheniformis; poly-gamma-glutamic acid; sodium nitrate; feeding fermentation; biomass

        1 材料與方法

        1.1 材料

        1.1.1 菌株 地衣芽孢桿菌WX-02,由華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物學(xué)國家重點(diǎn)實(shí)驗(yàn)室篩選和保藏,保藏號(hào)為CCTCC M208065。

        1.2 方法

        參考文獻(xiàn):

        [1] 陳詠竹,孫啟玲.γ-多聚谷氨酸的性質(zhì)、發(fā)酵生產(chǎn)及其應(yīng)用[J].微生物學(xué)通報(bào),2004,31(1): 122-126.

        [2] BAJAJ I, SINGHAL R. Poly(glutamic acid)- an emerging biopolymer of commercial interest[J]. Bioresour Technol,2011, 102(10):5551-5561.

        [3] 曹名鋒,金映虹,解 慧,等.γ-聚谷氨酸的微生物合成、相關(guān)基因及應(yīng)用展望[J].微生物學(xué)通報(bào),2011,38(3):388-395.

        [4] 蔡 謹(jǐn),孫章輝,王 雋,等.補(bǔ)料發(fā)酵工藝的應(yīng)用及其研究進(jìn)展[J].工業(yè)微生物,2005,35(1):42-48.

        [5] 呂 萌,梁金鐘,王風(fēng)青.補(bǔ)料發(fā)酵枯草芽孢桿菌合成γ-聚谷氨酸的研究[J].食品科學(xué),2011, 32(23):225-228.

        [6] 胡麗芳,李 欣,冀志霞,等.地衣芽胞桿菌WX-02補(bǔ)糖發(fā)酵聚γ-谷氨酸的工藝優(yōu)化[J]. 華中農(nóng)業(yè)大學(xué)學(xué)報(bào),2012,31(3):287-292.

        [7] YOON S H, DO J H, LEE S Y, et al. Production of poly-γ-glutamic acid by fed-batch culture of Bacillus licheniformis[J]. Biotechnology Letters,2000,22(7):585-588.

        [8] KRAFTA B, STROUSA M, TEGETMEYER H E. Microbial nitrate respiration -Genes, enzymes and environmental distribution[J]. Journal of Biotechnology, 2011,155(1):104-117.

        [9] CHENG C, ASADA Y, AIDA T. Production of γ-polyglutamic acid by Bacillus licheniformis A35 under denitrifying conditions[J]. Agricultural and Biological Chemistry,1989,53(9):2369-2375.

        [10] XU K D, STEWART P S, XIA F, et al. Spatial physiological heterogeneity in Pseudomonas aeruginosa biofilm is determined by oxygen availability[J]. Appl Environ Microbiol, 1998,64(10):4035-4039.

        [11] WEI X T, JI Z X, CHEN S W. Isolation of halotolerant Bacillus licheniformis WX-02 and regulatory effects of sodium chloride on yield and molecular sizes of poly-γ-glutamic acid[J]. Appl Biochem Biotechnol,2010,160(5):1332-1340.

        摘要:以地衣芽孢桿菌(Bacillus licheniformis)WX-02為研究對(duì)象,以提高其發(fā)酵產(chǎn)生的聚γ-谷氨酸的產(chǎn)量為研究目的,經(jīng)搖瓶發(fā)酵得到優(yōu)化的硝酸鈉補(bǔ)料方案,即初始硝酸鈉濃度為10 g/L,發(fā)酵至12 h時(shí)補(bǔ)加5 g/L硝酸鈉時(shí)聚γ-谷氨酸產(chǎn)量最大。將此補(bǔ)料方案在3 L發(fā)酵罐上進(jìn)行驗(yàn)證。結(jié)果表明,發(fā)酵得到的菌體生物量和聚γ-谷氨酸產(chǎn)量最大值分別為4.5 g/L和38.7 g/L,與對(duì)照相比分別提高55.2%和14.8%。通過補(bǔ)硝酸鈉可以提高菌體生物量和對(duì)谷氨酸的利用率,從而提高聚γ-谷氨酸的發(fā)酵水平。

        關(guān)鍵詞:地衣芽孢桿菌(Bacillus licheniformis);聚γ-谷氨酸;硝酸鈉;補(bǔ)料發(fā)酵;生物量

        中圖分類號(hào):TQ920.6 文獻(xiàn)標(biāo)識(shí)碼:A 文章編號(hào):0439-8114(2014)04-0903-04

        Optimizing Sodium Nitrate Fed-batch Culture on Poly(γ-glutamic acid) Fermentation by Bacillus licheniformis WX-02

        XU Shang-hua,HU Li-fang,CHEN Shou-wen

        (Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University , Wuhan 430070,China)

        Abstract: Bacillus licheniformis WX-02 was used to study the ways of improving poly(γ-glutamic acid)(γ-PGA) yield. The optimal conditions of sodium nitrate feeding for γ-PGA fermentation by Bacillus licheniformis WX-02 by flask-shaking fed-batch fermentation were the initial sodium nitrate concentration 10 g/L, and 5 g/L of sodium nitrate added when fermentation to 12 h. The fermentation was carried out in a 3 L fermentor, showing that the maximum of biomass and γ-PGA yield was 4.5 g/L and 38.7 g/L with increase of 55.2% and 14.8%, compared with the control. It is indicated that adding sodium nitrate can increase biomass, enhance the utilization of glutamate, and improve the γ-PGA fermentation.

        Key words: Bacillus licheniformis; poly-gamma-glutamic acid; sodium nitrate; feeding fermentation; biomass

        1 材料與方法

        1.1 材料

        1.1.1 菌株 地衣芽孢桿菌WX-02,由華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)微生物學(xué)國家重點(diǎn)實(shí)驗(yàn)室篩選和保藏,保藏號(hào)為CCTCC M208065。

        1.2 方法

        參考文獻(xiàn):

        [1] 陳詠竹,孫啟玲.γ-多聚谷氨酸的性質(zhì)、發(fā)酵生產(chǎn)及其應(yīng)用[J].微生物學(xué)通報(bào),2004,31(1): 122-126.

        [2] BAJAJ I, SINGHAL R. Poly(glutamic acid)- an emerging biopolymer of commercial interest[J]. Bioresour Technol,2011, 102(10):5551-5561.

        [3] 曹名鋒,金映虹,解 慧,等.γ-聚谷氨酸的微生物合成、相關(guān)基因及應(yīng)用展望[J].微生物學(xué)通報(bào),2011,38(3):388-395.

        [4] 蔡 謹(jǐn),孫章輝,王 雋,等.補(bǔ)料發(fā)酵工藝的應(yīng)用及其研究進(jìn)展[J].工業(yè)微生物,2005,35(1):42-48.

        [5] 呂 萌,梁金鐘,王風(fēng)青.補(bǔ)料發(fā)酵枯草芽孢桿菌合成γ-聚谷氨酸的研究[J].食品科學(xué),2011, 32(23):225-228.

        [6] 胡麗芳,李 欣,冀志霞,等.地衣芽胞桿菌WX-02補(bǔ)糖發(fā)酵聚γ-谷氨酸的工藝優(yōu)化[J]. 華中農(nóng)業(yè)大學(xué)學(xué)報(bào),2012,31(3):287-292.

        [7] YOON S H, DO J H, LEE S Y, et al. Production of poly-γ-glutamic acid by fed-batch culture of Bacillus licheniformis[J]. Biotechnology Letters,2000,22(7):585-588.

        [8] KRAFTA B, STROUSA M, TEGETMEYER H E. Microbial nitrate respiration -Genes, enzymes and environmental distribution[J]. Journal of Biotechnology, 2011,155(1):104-117.

        [9] CHENG C, ASADA Y, AIDA T. Production of γ-polyglutamic acid by Bacillus licheniformis A35 under denitrifying conditions[J]. Agricultural and Biological Chemistry,1989,53(9):2369-2375.

        [10] XU K D, STEWART P S, XIA F, et al. Spatial physiological heterogeneity in Pseudomonas aeruginosa biofilm is determined by oxygen availability[J]. Appl Environ Microbiol, 1998,64(10):4035-4039.

        [11] WEI X T, JI Z X, CHEN S W. Isolation of halotolerant Bacillus licheniformis WX-02 and regulatory effects of sodium chloride on yield and molecular sizes of poly-γ-glutamic acid[J]. Appl Biochem Biotechnol,2010,160(5):1332-1340.

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