氯化铵对紫色红曲霉固态发酵红曲色素和桔霉素合成的双向调控作用
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摘要
研究了氯化铵对紫色红曲霉M3103次级代谢产物中红曲色素和桔霉素合成代谢的影响及其调控机制。结果表明:在以红米为固态发酵基质的培养基中外加氯化铵能显著提高红曲色素产量,降低桔霉素产量;通过高效液相色谱-紫外可见光全波长扫描分析红曲色素组成,发现添加氯化铵显著提高了红曲黄色素和橙色素的产量;通过实时荧光定量PCR检测,红曲色素合成关键基因mppC、mppD、mppE、MpFasA2和MpPKS5的表达量与空白组相比均显著上调,而桔霉素合成关键基因ctnA和PksCT的表达量与空白组相比均显著下调。固态发酵中添加适量的氯化铵可影响紫色红曲霉M3103对营养物质吸收和代谢,有利于促进红曲色素尤其是黄色素的生物合成,抑制桔霉素的合成。
This study aimed to investigate the effect and regulatory mechanism of ammonium chloride on the production of Monascus pigments and citrinin in Monascus purpureus M3103.The results showed that the addition of ammonium chloride could significantly increase the yield of Monascus pigments but reduce the yield of citrinin in red rice as fermentation matrix.In particular,the production of Monascus yellow pigment and orange pigment were significantly enhanced by the addition of ammonium chloride through high performance liquid chromatography-ultraviolet and visible light(HPLC-UV-Vis) full wavelength scanner.Using quantitative real-time polymerase chain reaction,the relative expression of the key genes(mppC,mppD,mppE,MpFasA2 and MpPKS5) responsible for the production of Monascus pigments biosynthesis were detected significantly increased compared with the control group.However,the key genes(ctnA and PksCT) responsible for the production of Monascus citrinin biosynthesis were detected significantly decreased compared with the control group.Therefore,adding moderate amount of ammonium chloride to solid-state fermentation can affect the absorption and metabolism of nutrients by Monascus purpureus M3103.The addition of ammonium chloride is beneficial to promote the biosythesis of Monascus pigments,especially orange pigment,and inhibit the biosynthesis of citrinin.
This study aimed to investigate the effect and regulatory mechanism of ammonium chloride on the production of Monascus pigments and citrinin in Monascus purpureus M3103.The results showed that the addition of ammonium chloride could significantly increase the yield of Monascus pigments but reduce the yield of citrinin in red rice as fermentation matrix.In particular,the production of Monascus yellow pigment and orange pigment were significantly enhanced by the addition of ammonium chloride through high performance liquid chromatography-ultraviolet and visible light(HPLC-UV-Vis) full wavelength scanner.Using quantitative real-time polymerase chain reaction,the relative expression of the key genes(mppC,mppD,mppE,MpFasA2 and MpPKS5) responsible for the production of Monascus pigments biosynthesis were detected significantly increased compared with the control group.However,the key genes(ctnA and PksCT) responsible for the production of Monascus citrinin biosynthesis were detected significantly decreased compared with the control group.Therefore,adding moderate amount of ammonium chloride to solid-state fermentation can affect the absorption and metabolism of nutrients by Monascus purpureus M3103.The addition of ammonium chloride is beneficial to promote the biosythesis of Monascus pigments,especially orange pigment,and inhibit the biosynthesis of citrinin.
引文
[1]吕旭聪,翁星,韩妙坤,等.福建红曲中红曲菌的分离鉴定及菌株特性研究[J].中国食品学报,2012,12(2):88-97.
[2]郭季冬.红曲霉桔霉素合成相关基因的筛选与功能验证[D].天津:天津科技大学,2014.
[3]SHI Y C,LIAO V H,PAN T M.Monascin from red mold dioscorea as a novel antidiabetic and antioxidative stress agent in rats and Caenorhabditis elegans[J].Free Radical Biology&Medicine,2012,52(1):109-17.
[4]HSU L C,LIANG Y H,HSU Y W,et al.Antiinflammatory properties of yellow and orange pigments from Monascus purpureus NTU 568[J].Journal of Agricultural&Food Chemistry,2013,61(11):2796-2802.
[5]LEE C L,WEN J Y,HSU Y W,et al.Monascusfermented yellow pigments monascin and ankaflavin showed antiobesity effect via the suppression of differentiation and lipogenesis in obese rats fed a high-fat diet[J].Journal of Agricultural&Food Chemistry,2013,61(7):1493-500.
[6]HSU W H,CHEN T H,LEE B H,et al Monascin and ankaflavin act as natural AMPK activators with PPARαagonist activity to down-regulate nonalcoholic steatohepatitis in high-fat diet-fed C57BL/6 mice[J].Food&Chemical Toxicology,2014,64:94-103.
[7]SHI Y C,PAN T M.Characterization of a multifunctional Monascus isolate NTU 568 with high azaphilone pigments production[J].Food Biotechnology,2010,24(4):349-363.
[8]LEE C L,LIN P Y,HSU Y W,et al.Monascusfermented monascin and ankaflavin improve the memory and learning ability in amyloidβ-protein intracerebroventricular-infused rat via the suppression of Alzheimer’s disease risk factors[J].Journal of Functional Foods,2015,18:387-399.
[9]LADA R,ANA-MARIJA D,TIHANA Z G,et al Differential activation of MAPKs by individual and combined ochratoxin A and citrinin treatments in porcine kidney PK15 cells[J].Toxicon,2014,90:174-83.
[10]LEE B H,PAN T M.Benefit of Monascus-fermented products for hypertension prevention:a review[J]Applied Microbiology&Biotechnology,2012,94(5):1151-1161.
[11]YANG J,CHEN Q,WANG W,et al.Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation[J].Journal of Bioscience&Bioengineering,2015,119(5):564-569.
[12]WANG L,DAI Y,CHEN W,et al.Effects of light intensity and color on the biomass,extracellular red pigment,and citrinin production of Monascus ruber[J].Journal of Agricultural&Food Chemistry,2016,64(50):9506-9514.
[13]LV J,ZHANG B B,LIU X D,et al.Enhanced production of natural yellow pigments from Monascus purpureus by liquid culture:The relationship between fermentation conditions and mycelial morphology[J].Journal of Bioscience&Bioengineering,2017,124(4):452-458.
[14]梁斐,杜欣军,张燕,等.红曲霉固态发酵中桔霉素的抑制研究[J].食品科技,2016,41(3):2-6.
[15]SHI K,SONG D,CHEN G,et al.Controlling composition and color characteristics of Monascus pigments by pH and nitrogen sources in submerged fermentation[J].Journal of Bioscience&Bioengineering,2015,120(2):145-154.
[16]童爱均,吕旭聪,刘晓艳,等.固体发酵基质和外加碳氮源对红曲霉色素产量的影响[J].中国调味品,2015,40(5):5-10.
[17]WANG C,DI C,CHEN M,et al.Stimulatory effects of blue light on the growth,monascin and ankaflavin production in Monascus[J].Biotechnology Letters,2015,37(5):1043.
[18]岳建明,杨强,肖潇,等.铵盐对紫色红曲霉合成代谢红曲色素及桔霉素的影响[J].食品科学,2016,37(5):102-107.
[19]BALAKRISHNAN B,KARKI S,CHIU S H,et al Genetic localization and in vivo characterization of a Monascus azaphilone pigment biosynthetic gene cluster[J].Applied Microbiology&Biotechnology,2013,97(14):6337-6345.
[20]BALAKRISHNAN B,CHEN C C,PAN T M,et al.Mpp7 controls regioselective Knoevenagel condensation during the biosynthesis of Monascus azaphilone pigments[J].Tetrahedron Letters,2014,55(9):1640-1643.
[21]CHEN Y P,YUAN G F,HSIEH S Y,et al.I-dentification of the mokH gene encoding transcription factor for the upregulation of monacolin Kbiosynthesis in monascus pilosus[J].Journal of Agricultural&Food Chemistry,2010,58(1):287-293.
[22]LI Y P,XU Y,HUANG Z B.Isolation and characterization of the citrinin biosynthetic gene cluster from Monascus aurantiacus[J].Biotechnology Letters,2012,34(1):131-136.
[23]杨宁,郭红珍,郝林.山西老陈醋大曲红曲霉分离鉴定及生物学特性研究[J].山西农业大学学报(自然科学版),2006,26(1):4-6.
[24]LIU Q,XIE N,YI H,et al.MpigE,a gene involved in pigment biosynthesis in Monascus ruber,M7[J].Applied Microbiology&Biotechnology,2014,98(1):285-296.
[25]BALAKRISHNAN B,CHANDRAN R,PARK S H,et al.A new protein factor in the product formation of non-reducing fungal polyketide synthase with a c-terminus reductive domain[J].Journal of Microbiology&Biotechnology,2015,25(10):1648-1652.
[26]LIN C H,LIN T H,PAN T M.Alleviation of metabolic syndrome by monascin and ankaflavin:the perspective of Monascus functional foods[J].Food&Function,2017,8(6):2102-2109.
[2]郭季冬.红曲霉桔霉素合成相关基因的筛选与功能验证[D].天津:天津科技大学,2014.
[3]SHI Y C,LIAO V H,PAN T M.Monascin from red mold dioscorea as a novel antidiabetic and antioxidative stress agent in rats and Caenorhabditis elegans[J].Free Radical Biology&Medicine,2012,52(1):109-17.
[4]HSU L C,LIANG Y H,HSU Y W,et al.Antiinflammatory properties of yellow and orange pigments from Monascus purpureus NTU 568[J].Journal of Agricultural&Food Chemistry,2013,61(11):2796-2802.
[5]LEE C L,WEN J Y,HSU Y W,et al.Monascusfermented yellow pigments monascin and ankaflavin showed antiobesity effect via the suppression of differentiation and lipogenesis in obese rats fed a high-fat diet[J].Journal of Agricultural&Food Chemistry,2013,61(7):1493-500.
[6]HSU W H,CHEN T H,LEE B H,et al Monascin and ankaflavin act as natural AMPK activators with PPARαagonist activity to down-regulate nonalcoholic steatohepatitis in high-fat diet-fed C57BL/6 mice[J].Food&Chemical Toxicology,2014,64:94-103.
[7]SHI Y C,PAN T M.Characterization of a multifunctional Monascus isolate NTU 568 with high azaphilone pigments production[J].Food Biotechnology,2010,24(4):349-363.
[8]LEE C L,LIN P Y,HSU Y W,et al.Monascusfermented monascin and ankaflavin improve the memory and learning ability in amyloidβ-protein intracerebroventricular-infused rat via the suppression of Alzheimer’s disease risk factors[J].Journal of Functional Foods,2015,18:387-399.
[9]LADA R,ANA-MARIJA D,TIHANA Z G,et al Differential activation of MAPKs by individual and combined ochratoxin A and citrinin treatments in porcine kidney PK15 cells[J].Toxicon,2014,90:174-83.
[10]LEE B H,PAN T M.Benefit of Monascus-fermented products for hypertension prevention:a review[J]Applied Microbiology&Biotechnology,2012,94(5):1151-1161.
[11]YANG J,CHEN Q,WANG W,et al.Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation[J].Journal of Bioscience&Bioengineering,2015,119(5):564-569.
[12]WANG L,DAI Y,CHEN W,et al.Effects of light intensity and color on the biomass,extracellular red pigment,and citrinin production of Monascus ruber[J].Journal of Agricultural&Food Chemistry,2016,64(50):9506-9514.
[13]LV J,ZHANG B B,LIU X D,et al.Enhanced production of natural yellow pigments from Monascus purpureus by liquid culture:The relationship between fermentation conditions and mycelial morphology[J].Journal of Bioscience&Bioengineering,2017,124(4):452-458.
[14]梁斐,杜欣军,张燕,等.红曲霉固态发酵中桔霉素的抑制研究[J].食品科技,2016,41(3):2-6.
[15]SHI K,SONG D,CHEN G,et al.Controlling composition and color characteristics of Monascus pigments by pH and nitrogen sources in submerged fermentation[J].Journal of Bioscience&Bioengineering,2015,120(2):145-154.
[16]童爱均,吕旭聪,刘晓艳,等.固体发酵基质和外加碳氮源对红曲霉色素产量的影响[J].中国调味品,2015,40(5):5-10.
[17]WANG C,DI C,CHEN M,et al.Stimulatory effects of blue light on the growth,monascin and ankaflavin production in Monascus[J].Biotechnology Letters,2015,37(5):1043.
[18]岳建明,杨强,肖潇,等.铵盐对紫色红曲霉合成代谢红曲色素及桔霉素的影响[J].食品科学,2016,37(5):102-107.
[19]BALAKRISHNAN B,KARKI S,CHIU S H,et al Genetic localization and in vivo characterization of a Monascus azaphilone pigment biosynthetic gene cluster[J].Applied Microbiology&Biotechnology,2013,97(14):6337-6345.
[20]BALAKRISHNAN B,CHEN C C,PAN T M,et al.Mpp7 controls regioselective Knoevenagel condensation during the biosynthesis of Monascus azaphilone pigments[J].Tetrahedron Letters,2014,55(9):1640-1643.
[21]CHEN Y P,YUAN G F,HSIEH S Y,et al.I-dentification of the mokH gene encoding transcription factor for the upregulation of monacolin Kbiosynthesis in monascus pilosus[J].Journal of Agricultural&Food Chemistry,2010,58(1):287-293.
[22]LI Y P,XU Y,HUANG Z B.Isolation and characterization of the citrinin biosynthetic gene cluster from Monascus aurantiacus[J].Biotechnology Letters,2012,34(1):131-136.
[23]杨宁,郭红珍,郝林.山西老陈醋大曲红曲霉分离鉴定及生物学特性研究[J].山西农业大学学报(自然科学版),2006,26(1):4-6.
[24]LIU Q,XIE N,YI H,et al.MpigE,a gene involved in pigment biosynthesis in Monascus ruber,M7[J].Applied Microbiology&Biotechnology,2014,98(1):285-296.
[25]BALAKRISHNAN B,CHANDRAN R,PARK S H,et al.A new protein factor in the product formation of non-reducing fungal polyketide synthase with a c-terminus reductive domain[J].Journal of Microbiology&Biotechnology,2015,25(10):1648-1652.
[26]LIN C H,LIN T H,PAN T M.Alleviation of metabolic syndrome by monascin and ankaflavin:the perspective of Monascus functional foods[J].Food&Function,2017,8(6):2102-2109.
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