产紫青霉突变菌株Li-3-9的发酵条件优化及红色素安全性评价
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摘要
本实验对前期研究得到的具有高产红色素能力的产紫青霉突变菌株Penicilliumpurpurogenum Li-3-9的发酵条件进行优化的同时作出了对红色素的安全性评价。在单因素试验的基础上,采用Box-Behnken试验设计优化了蔗糖浓度、酵母膏浓度和装液量对红色素色价的影响;并且研究了P. purpurogenum Li-3-9产生的红色素对斑马鱼胚胎的毒性效应。结果表明,最佳发酵条件为:蔗糖40g/L、酵母膏4g/L、装液量为45mL、接种量4%、培养温度32℃、初始pH值6.9、培养时间168h、摇床转速150r/min,红色素色价最高达到了11.4U/mL。通过计算斑马鱼胚胎致死率、心率、畸形率及孵化率可知,该色素无致死毒性,并且对斑马鱼胚胎发育具有一定的促进作用。
The fermentation conditions and red pigment safety of Penicillium purpurogenum Li-3-9 were investigated. On the basis of single factor test, the Box-Behnken was used to study the effect of concentration of sucrose and yeast extract, and liquid volume on the red color value. The toxic effect of red pigment on zebrafish embryos was also studied. The results indicated that the optimum fermentation conditions were as follows: culture medium including sucrose 40 g/L and yeast extract 4 g/L in liquid volume of 50 mL, inoculation guantity of 4%, incubation under the temperature of 32°C at pH 6.9 in duration of 168 h at rotation speed of150 r/min. Under such conditions the color value reached 11.4 U/mL. Results of calculation of the zebrafish embryo mortality, heart rate, malformation rate and hatching rate indicated that the red pigment was non-toxic and no serious malformation occurred. On the contrary, it has promoting effect on zebrafish embryo development to a certain extent.
The fermentation conditions and red pigment safety of Penicillium purpurogenum Li-3-9 were investigated. On the basis of single factor test, the Box-Behnken was used to study the effect of concentration of sucrose and yeast extract, and liquid volume on the red color value. The toxic effect of red pigment on zebrafish embryos was also studied. The results indicated that the optimum fermentation conditions were as follows: culture medium including sucrose 40 g/L and yeast extract 4 g/L in liquid volume of 50 mL, inoculation guantity of 4%, incubation under the temperature of 32°C at pH 6.9 in duration of 168 h at rotation speed of150 r/min. Under such conditions the color value reached 11.4 U/mL. Results of calculation of the zebrafish embryo mortality, heart rate, malformation rate and hatching rate indicated that the red pigment was non-toxic and no serious malformation occurred. On the contrary, it has promoting effect on zebrafish embryo development to a certain extent.
引文
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Cao H,Ye H,Li C,Zheng LL,Li Y,Ouyang QF,2014.Effect of microencapsulated cell preparation technology and conditions on the catalytic performance of Penicillium purpurogenum Li-3 strain cells.Process Biochemistry,49(5):791-796
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Chai YJ,Cui CB,Li CW,Wu CJ,Tian CK,Hja W,2012.Activation of the dormant secondary metabolite production by introducing gentamicin-resistance in a marine-derived Penicillium purpurogenum G59.Marine Drugs,10(3):559-582
Chulyoung K,Heeyong J,Yong OK,Chul SS,2006.Antimicrobial activities of amino acid derivatives of monascus pigments.FEMS Microbiology Letters,264(1):117-124
Feng Y,Shao Y,Chen F,2012.Monascus pigments.Applied Microbiology and Biotechnology,96(6):1421-1440
He QX,Dong ZL,Chu J,Sun GJ,Han LW,Han J,Liu KC,2015.Toxicity study of aloe-emodin on embryonic development and motor behavior of zebrafish.Shandong Science,3:23-28(in Chinese)
Hsu WH,Pan TM,2012.Monascus purpureus-fermented products and oral cancer:a review.Applied Microbiology and Biotechnology,93(5):1831-1842
Lee KJ,Nallathamby PD,Browning LM,Osgood CJ,Xu XHN,2007.In vivo imaging of transport and biocompatibility of single silver nanoparticles in early development of zebrafish embryos.ACS Nano,1(2):133-143
Li JB,Jia W,Zhao QS,2014.Excessive nitrite affects zebrafish valvulogenesis through yielding too much NO signaling.PLoS One,9(3):27-28
Niu YJ,Cao H,Huang XL,Tong JM,Shui JH,Yang YX,2017.Breeding of high-yield red pigment by P.purpurogenum Li-3 and its biological activity.China Brewing,36(3):30-34(in Chinese)
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Mapari SAS,Thrane U,Meyer AS,2010.Fungal polyketide azaphilone pigments as future natural food colorants.Trends in Biotechnology,28(6):300-307
Kang B,Zhang X,Wu Z,Wang ZL,Park S,2014.Production of citrinin-free Monascus pigments by submerged culture at low pH.Enzyme and Microbial Technology,55:50-57
Kongruang S,2011.Growth kinetics of biopigment production by Thai isolated Monascus purpureus in a stirred tank bioreactor.Journal of Industrial Microbiology&Biotechnology,38(1):93-98
Papagianni M,2004.Fungal morphology and metabolite production in submerged mycelial processes.Biotechnology Advances,22(3):189-259
Santos-Ebinuma VC,Roberto IC,Teixeira MFS,Pessoa A,2013.Improving of red colorants production by a new P.purpurogenum strain in submerged culture and the effect of different parameters in their stability.Biotechnology Progress,29(3):778-785
Singh D,Kaur G,2014.Swainsonine,a novel fungal metabolite:optimization of fermentative production and bioreactor operations using evolutionary programming.Bioprocess and Biosystems Engineering,37(8):1599-1607
Tuli HS,Chaudhary P,Beniwal V,Sharma AK,2015.Microbial pigments as natural color sources:current trends and future perspectives.Journal of Food Science and Technology,52(8):1-10
Velmurugan P,Lee YH,Venil CK,Lakshmanaperumalsamy P,Chae JC,Oh BT,2010.Effect of light on growth,intracellular and extracellular pigment production by five pigment-producing filamentous fungi in synthetic medium.Journal of Bioscience&Bioengineering,109(4):346-350
Wang JJ,Xu C,Tu YJ,Fu ZW,Liu WP,2007.Advances in experimental research and application of zebrafish and its embryos in toxicology.Asian Journal of Ecotoxicology,2(2):123-135(in Chinese)
Wang FQ,Xu B,Sun YF,Zang JY,Li XM,Wang NF,2013.Identification of an antarctic fungus and analysis of its secreted pigment properties.Chinese Journal of Food Science,3:189-195(in Chinese)
Yang J,Chen Q,Wang W,Hu JJ,Hu C,2015.Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation.Journal of Bioscience and Bioengineering,119(5):564-569
Yin JS,2012.Breeding of rhizoma polysaccharide and functional red yeast and optimization of production process.Master’s Thesis,Shandong Agricultural University,Qingdao.24-29(in Chinese)
Zhang X,Cao H,Li C,Niu YJ,Yang L,2016.Study on fermentation conditions,pigment stability and safety of Penicillium purpurogenum.China Food Additives,2:68-74(in Chinese)
Zhou QX,Yang L,Wang CX,Xue Y,Xue CH,Xu J,2016.Physicochemical properties of a sea urchin co-added bacteria and its fermentation product yellow pigment.Chinese Journal of Food Science,12:240-246(in Chinese)
Zhu HJ,2013.Study on the mechanism of the directional regulator of safe pigment-yielding red yeast.Master Thesis,Hubei University of Technology,Wuhan.18-21(in Chinese)
蔡道基,杨佩芝,龚瑞忠,1989.化学农药环境安全评价试验准则.北京:国家环境保护局.5-79
何秋霞,董贞兰,楚杰,孙桂金,韩利文,韩健,刘可春,2015.芦荟大黄素对斑马鱼胚胎发育及运动行为学的毒性研究.山东科学,3:23-28
牛玉静,曹红,黄晓林,童军茂,隋佳慧,杨雨鑫,2017.复合诱变P.purpurogenum Li-3选育高产红色素及其生物活性研究.中国酿造,36(3):30-34
王佳佳,徐超,屠云杰,傅正伟,刘维屏,2007.斑马鱼及其胚胎在毒理学中的实验研究与应用进展.生态毒理学报,2(2):123-135
王风芹,许宾,孙云飞,臧家业,李雪梅,王能飞,2013.1株南极真菌的鉴定及其分泌色素性质分析.中国食品学报,3:189-195
殷军帅,2012.红曲色素和功能性红曲的菌种选育和生产工艺的优化.山东农业大学硕士学位论文,青岛.24-29
朱宏军,2013.安全型色素红曲的定向调节剂其机理的研究.湖北工业大学硕士学位论文,武汉.18-21
张馨,曹红,李春,牛玉静,杨露,2016.产紫青霉菌的发酵条件及色素稳定性、安全性的研究.中国食品添加剂,2:68-74
周庆新,杨鲁,王昕岑,薛勇,薛长湖,徐杰,2016.一株海胆共附菌及其发酵产物黄色素的理化性质分析.中国食品学报,12:240-246
Cao H,Ye H,Li C,Zheng LL,Li Y,Ouyang QF,2014.Effect of microencapsulated cell preparation technology and conditions on the catalytic performance of Penicillium purpurogenum Li-3 strain cells.Process Biochemistry,49(5):791-796
Chai DJ,Yang PZ,Gong RZ,1989.Chemical pesticide environmental safety evaluation test criteria.National Environmental Protection Agency,Beijing.75-79(in Chinese)
Chai YJ,Cui CB,Li CW,Wu CJ,Tian CK,Hja W,2012.Activation of the dormant secondary metabolite production by introducing gentamicin-resistance in a marine-derived Penicillium purpurogenum G59.Marine Drugs,10(3):559-582
Chulyoung K,Heeyong J,Yong OK,Chul SS,2006.Antimicrobial activities of amino acid derivatives of monascus pigments.FEMS Microbiology Letters,264(1):117-124
Feng Y,Shao Y,Chen F,2012.Monascus pigments.Applied Microbiology and Biotechnology,96(6):1421-1440
He QX,Dong ZL,Chu J,Sun GJ,Han LW,Han J,Liu KC,2015.Toxicity study of aloe-emodin on embryonic development and motor behavior of zebrafish.Shandong Science,3:23-28(in Chinese)
Hsu WH,Pan TM,2012.Monascus purpureus-fermented products and oral cancer:a review.Applied Microbiology and Biotechnology,93(5):1831-1842
Lee KJ,Nallathamby PD,Browning LM,Osgood CJ,Xu XHN,2007.In vivo imaging of transport and biocompatibility of single silver nanoparticles in early development of zebrafish embryos.ACS Nano,1(2):133-143
Li JB,Jia W,Zhao QS,2014.Excessive nitrite affects zebrafish valvulogenesis through yielding too much NO signaling.PLoS One,9(3):27-28
Niu YJ,Cao H,Huang XL,Tong JM,Shui JH,Yang YX,2017.Breeding of high-yield red pigment by P.purpurogenum Li-3 and its biological activity.China Brewing,36(3):30-34(in Chinese)
Mapari SAS,Hansen ME,Meyer AS,Thrane U,2008.Computerized screening for novel producers of Monascus-like food pigments in Penicillium species.Journal of Agricultural&Food Chemistry,56(21):9981-9989
Mapari SAS,Thrane U,Meyer AS,2010.Fungal polyketide azaphilone pigments as future natural food colorants.Trends in Biotechnology,28(6):300-307
Kang B,Zhang X,Wu Z,Wang ZL,Park S,2014.Production of citrinin-free Monascus pigments by submerged culture at low pH.Enzyme and Microbial Technology,55:50-57
Kongruang S,2011.Growth kinetics of biopigment production by Thai isolated Monascus purpureus in a stirred tank bioreactor.Journal of Industrial Microbiology&Biotechnology,38(1):93-98
Papagianni M,2004.Fungal morphology and metabolite production in submerged mycelial processes.Biotechnology Advances,22(3):189-259
Santos-Ebinuma VC,Roberto IC,Teixeira MFS,Pessoa A,2013.Improving of red colorants production by a new P.purpurogenum strain in submerged culture and the effect of different parameters in their stability.Biotechnology Progress,29(3):778-785
Singh D,Kaur G,2014.Swainsonine,a novel fungal metabolite:optimization of fermentative production and bioreactor operations using evolutionary programming.Bioprocess and Biosystems Engineering,37(8):1599-1607
Tuli HS,Chaudhary P,Beniwal V,Sharma AK,2015.Microbial pigments as natural color sources:current trends and future perspectives.Journal of Food Science and Technology,52(8):1-10
Velmurugan P,Lee YH,Venil CK,Lakshmanaperumalsamy P,Chae JC,Oh BT,2010.Effect of light on growth,intracellular and extracellular pigment production by five pigment-producing filamentous fungi in synthetic medium.Journal of Bioscience&Bioengineering,109(4):346-350
Wang JJ,Xu C,Tu YJ,Fu ZW,Liu WP,2007.Advances in experimental research and application of zebrafish and its embryos in toxicology.Asian Journal of Ecotoxicology,2(2):123-135(in Chinese)
Wang FQ,Xu B,Sun YF,Zang JY,Li XM,Wang NF,2013.Identification of an antarctic fungus and analysis of its secreted pigment properties.Chinese Journal of Food Science,3:189-195(in Chinese)
Yang J,Chen Q,Wang W,Hu JJ,Hu C,2015.Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation.Journal of Bioscience and Bioengineering,119(5):564-569
Yin JS,2012.Breeding of rhizoma polysaccharide and functional red yeast and optimization of production process.Master’s Thesis,Shandong Agricultural University,Qingdao.24-29(in Chinese)
Zhang X,Cao H,Li C,Niu YJ,Yang L,2016.Study on fermentation conditions,pigment stability and safety of Penicillium purpurogenum.China Food Additives,2:68-74(in Chinese)
Zhou QX,Yang L,Wang CX,Xue Y,Xue CH,Xu J,2016.Physicochemical properties of a sea urchin co-added bacteria and its fermentation product yellow pigment.Chinese Journal of Food Science,12:240-246(in Chinese)
Zhu HJ,2013.Study on the mechanism of the directional regulator of safe pigment-yielding red yeast.Master Thesis,Hubei University of Technology,Wuhan.18-21(in Chinese)
蔡道基,杨佩芝,龚瑞忠,1989.化学农药环境安全评价试验准则.北京:国家环境保护局.5-79
何秋霞,董贞兰,楚杰,孙桂金,韩利文,韩健,刘可春,2015.芦荟大黄素对斑马鱼胚胎发育及运动行为学的毒性研究.山东科学,3:23-28
牛玉静,曹红,黄晓林,童军茂,隋佳慧,杨雨鑫,2017.复合诱变P.purpurogenum Li-3选育高产红色素及其生物活性研究.中国酿造,36(3):30-34
王佳佳,徐超,屠云杰,傅正伟,刘维屏,2007.斑马鱼及其胚胎在毒理学中的实验研究与应用进展.生态毒理学报,2(2):123-135
王风芹,许宾,孙云飞,臧家业,李雪梅,王能飞,2013.1株南极真菌的鉴定及其分泌色素性质分析.中国食品学报,3:189-195
殷军帅,2012.红曲色素和功能性红曲的菌种选育和生产工艺的优化.山东农业大学硕士学位论文,青岛.24-29
朱宏军,2013.安全型色素红曲的定向调节剂其机理的研究.湖北工业大学硕士学位论文,武汉.18-21
张馨,曹红,李春,牛玉静,杨露,2016.产紫青霉菌的发酵条件及色素稳定性、安全性的研究.中国食品添加剂,2:68-74
周庆新,杨鲁,王昕岑,薛勇,薛长湖,徐杰,2016.一株海胆共附菌及其发酵产物黄色素的理化性质分析.中国食品学报,12:240-246