褪黑素诱导糖蜜废醪液培养单针藻Monoraphidium sp.FXY-10产生物燃料
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摘要
研究褪黑素对糖蜜酒精废醪液中单针藻Monoraphidium sp. FXY-10生长和油脂积累的影响。结果表明,以糖蜜酒精废醪液为基础培养基,添加10~(-3)μmol/L的褪黑素,微藻中的油脂含量达到68.69%,是对照组的1.15倍,且最高生物量为1.22g/L,相比对照组提高了41.86%。与对照组相比,藻细胞内蛋白质和碳水化合物含量分别是对照组的0.61和1.41倍。在微藻培养过程中,糖蜜废醪液中的COD、总氮及总磷的清除率分别达到92.33%、90.07%和86.04%。此外,外源褪黑素的添加提高了乙酰辅酶A羧化酶(acetyl coenzyme A carboxylase,ACCase)、苹果酸酶(malic enzyme,ME)的活性,而降低了磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)的活性。初步技术经济分析表明,1kg单针藻Monoraphidium sp. FXY-10的干燥粉可以制备535.8g生物柴油。研究表明,褪黑素可以促进糖蜜酒精废醪液中微藻的生长和油脂的积累,降低培养成本,为微藻扩大化生产提供了新的理论基础。
The effects of melatonin on the biomass and lipid production in Monoraphidium sp. FXY-10 under molasses wastewater were studied. The molasses wastewater was used as a growth media for the cultivation of Monoraphidium sp. FXY-10. Lipid content of algae was reached 68.69 % with 10~(-3) μmol/L melatonin-treated, which was increased by 1.15-fold. The biomass was 1.22 g/L, representing 41.86%higher than that of single molasses wastewater. And the protein and carbohydrate contents were increased and decreased by 1.41-and 0.61-fold, respectively. The removal rate of COD, total nitrogen and total phosphorus in molasses waste mash reached 92.33%, 90.07% and 86.04%, respectively in the microalgae cultivation. Furthermore, the lipid biosynthesis-related enzyme activities correlated with the increased lipid accumulation. The ACCase, and ME activities were considerably up-regulated with PEPC activity but were significantly down-regulated by melatonin induction. The yield analysis indicated that 1 kg dried biomass of Monoraphidium sp. FXY-10 might produce about 535.8 g of biodiesel. These low cost findings demonstrated that exogenous melatonin could promote the biomass and lipid production in microalgae under molasses wastewater, and provide a theoretical basis for the large-scale microalgae cultivation.
The effects of melatonin on the biomass and lipid production in Monoraphidium sp. FXY-10 under molasses wastewater were studied. The molasses wastewater was used as a growth media for the cultivation of Monoraphidium sp. FXY-10. Lipid content of algae was reached 68.69 % with 10~(-3) μmol/L melatonin-treated, which was increased by 1.15-fold. The biomass was 1.22 g/L, representing 41.86%higher than that of single molasses wastewater. And the protein and carbohydrate contents were increased and decreased by 1.41-and 0.61-fold, respectively. The removal rate of COD, total nitrogen and total phosphorus in molasses waste mash reached 92.33%, 90.07% and 86.04%, respectively in the microalgae cultivation. Furthermore, the lipid biosynthesis-related enzyme activities correlated with the increased lipid accumulation. The ACCase, and ME activities were considerably up-regulated with PEPC activity but were significantly down-regulated by melatonin induction. The yield analysis indicated that 1 kg dried biomass of Monoraphidium sp. FXY-10 might produce about 535.8 g of biodiesel. These low cost findings demonstrated that exogenous melatonin could promote the biomass and lipid production in microalgae under molasses wastewater, and provide a theoretical basis for the large-scale microalgae cultivation.
引文
[1]CHATTOPADHYAY S,KAREMORE A,DAS S,et al.Biocatalytic production of biodiesel from cottonseed oil:standardization of process parameters and comparison of fuel characteristics[J].Applied Energy,2011,88(4):1251-1256.
[2]WU X,LEUNG D Y C.Optimization of biodiesel production from camelina oil using orthogonal experiment[J].Applied Energy,2011,88(11):3615-3624.
[3]李川,胡献国.绿色生物质灰催化剂用于合成生物柴油[J].石油学报(石油加工),2018,34(1):78-85.LI Chuan,HU Xianguo.Green biomass ash catalyst for biodiesel synthesis[J].Journal of Petroleum(Petroleum Processing),2018,34(1):78-85.
[4]王常文,崔方方,宋宇.生物柴油的研究现状及发展前景[J].中国油脂,2014,39(5):44-48.WANG Changwen,CUI Fangfang,SONG Yu.Research situation and development prospect of biodiesel[J].Chinese Oils and Fats,2014,39(5):44-48.
[5]STEPHENSON P G,MOORE C M,TERRY M J,et al.Improving photosynthesis for algal biofuels:toward a green revolution[J].Trends in Biotechnology,2011,29(12):615-623.
[6]CARIOCA J O,HILUY FILHO J J,LEAL M R,et al.The hard choice for alternative biofuels to diesel in Brazil[J].Biotechnology Advances,2009,27(6):1043-1050.
[7]郭丹,银建中.微藻制备生物柴油的技术进展[J].化工装备技术,2014,35(4):4-9.GUO Dan,YIN Jianzhong.Technology progress of biodiesel production from microalgae[J].Chemical Equipment Technology,2014,35(4):4-9.
[8]方正,吕德义.微藻制备生物柴油的研究进展[J].现代化工,2017,37(9):57-61.FANG Zheng,LüDeyi.Research progress on biodiesel production by microalgae[J].Modern Chemical Industry,2017,37(9):57-61.
[9]李斐,白净,常春,等.微藻生物柴油成本控制技术进展[J].现代化工,2015,35(5):16-20.LI Fei,BAI Jing,CHANG Chun,et al.Developments of cost control technologies to produce biodiesel from microalgae[J].Modern Chemical Industry,2015,35(5):16-20.
[10]NAYAK M,KAREMORE A,SEN R.Performance evaluation of microalgae for concomitant wastewater bioremediation,CO2,biofixation and lipid biosynthesis for biodiesel application[J].Algal Research,2016,16:216-223.
[11]FERNáNDEZLINARES L C,GUERRERO B C,DURáN P E,et al.Assessment of Chlorella vulgaris and indigenous microalgae biomass with treated wastewater as growth culture medium[J].Bioresource Technology,2017,244(Pt 1):400-406.
[12]周祖光.糖蜜酒精生产废醪液资源化利用探析[J].环境科学与技术,2005,28(2):98-100.ZHOU Zuguang.Research on the utilization of molasses waste water[J].Environmental Science and Technology,2005,28(2):98-100.
[13]张跃彬,高正卿.蔗糖副产物滤泥、糖蜜酒精废醪液的开发利用[J].中国糖料,2009(1):63-64.ZHANG Yuebin,GAO Zhengqing.Development and utilization on filter mud of cane sugar by-product and wastewater from alcoholic fermentation of molasses[J].Sugar Crops of China,2009(1):63-64.
[14]CHAO M,WEN H,XING D,et al.Molasses wastewater treatment and lipid production at low temperature conditions by a microalgal mutant Scenedesmus sp.Z-4[J].Biotechnology for Biofuels,2017,10(1):111-123.
[15]YU Z,PEI H Y,JIANG L Q,et al.Phytohormone addition coupled with nitrogen depletion almost tripled the lipid productivities in two algae[J].Bioresource Technology,2018,247:904-914.
[16]ZHANG H,ZHANG N,YANG R,et al.Melatonin promotes seed germination under high salinity by regulating antioxidant systems,ABA and GA4 interaction in cucumber(Cucumis sativus L.)[J].Journal of Pineal Research,2015,57(3):269-279.
[17]BYEON Y,BACK K.An increase in melatonin in transgenic rice causes pleiotropic phenotypes,including enhanced seedling growth,delayed flowering,and low grain yield[J].Journal of Pineal Research,2014,56(4):408-414.
[18]NAWAZ M A,JIAO Y,CHEN C,et al.Melatonin pretreatment improves vanadium stress tolerance of watermelon seedlings by reducing vanadium concentration in the leaves and regulating melatonin biosynthesis and antioxidant-related gene expression[J].Journal of Plant Physiology,2018,220:115-127.
[19]YU X,ZHAO P,HE C,et al.Isolation of a novel strain of Monoraphidium sp and characterization of its potential application as biodiesel feedstock[J].Bioresource Technology,2012,121(7):256-262.
[20]CHE R,HUANG L,XU J W,et al.Effect of fulvic acid induction on the physiology,metabolism,and lipid biosynthesis-related gene transcription of Monoraphidium sp.FXY-10[J].Bioresource Technology,2016,227:324-334.
[21]BERGES J A,FISHER A E,HARRISON P J.A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana[J].Marine Biology,1993,115(2):187-193.
[22]ZHAO Y,LI D,DING K,et al.Production of biomass and lipids by the oleaginous microalgae Monoraphidium sp.QLY-1 through heterotrophic cultivation and photo-chemical modulator induction[J].Bioresour Technol,2016,211:669-676.
[23]ZHAO Y,LI D,XU J W,et al.Melatonin enhances lipid production in Monoraphidium sp.QLY-1 under nitrogen deficiency conditions via a multi-level mechanism[J].Bioresource Technology,2018,259:46-35.
[24]WEI W,LI Q T,CHU Y N,et al.Melatonin enhances plant growth and abiotic stress tolerance in soybean plants[J].Journal of Experimental Botany,2015,66(3):695-707.
[25]SHI L X,THEG S M.The chloroplast protein import system:from algae to trees[J].Biochim Biophys Acta,2013,1833(2):314-331.
[26]金永华,周伟能,吴玉林.乙酰辅酶A羧化酶抑制剂的研究进展[J].海峡药学,2018,30(1):9-14.JIN Yonghua,ZHOU Weineng,WU Yulin.Progress in the research of acetyl coenzyme a carboxylase inhibitors[J].Strait Pharmaceutical Journal,2018,30(1):9-14.
[27]GOODENOUG H U,BLABY I,CASERO D,et al.The path to triacylglyceride obesity in the sta6 strain of Chlamydomonas reinhardtii[J].Eukaryotic Cell,2014,13(5):591-613.
[28]MA X,LIU J,LIU B,et al.Physiological and biochemical changes reveal stress-associated photosynthetic carbon partitioning into triacylglycerol in the oleaginous marine alga Nannochloropsis oculata[J].Algal Research,2016,16:28-35
[29]吕娜娜,朱葆华,鹿琳,等.过表达三角褐指藻苹果酸酶基因提高E.coli脂肪酸合成能力研究[J].中国海洋大学学报(自然科学版),2016,46(5):65-69.LüNana,ZHU Baohua,LU Lin,et al.Overexpression of malic enzyme gene from phaeodactylum tricornutum promotes fatty acids production in Escherichia coli[J].Periodical of Ocean University of China(Natural Science Edition),2016,46(5):65-69.
[30]COURCHESNE N,PARISIEN A,WANG B,et al.Enhancement of lipid production using biochemical,genetic and transcription factor engineering approaches[J].Journal of Biotechnology,2009,142(1):31-41.
[31]霍垲,陆巍,李霞.干旱胁迫下调节ATP的含量对提高转玉米C4型pepc水稻光合速率的影响[J].中国生态农业学报,2015,23(5):605-613.HUO Kai,LU Wei,LI Xia.Effect of regulating ATP on improving photosynthetic rate of transgenic rice with overexpressing maize C4pepc under drought stress[J].Chinese Journal of Eco-Agriculture,2015,23(5):605-613.
[32]DENG X,CAI J,LI Y,et al.Expression and knockdown of the PEPC1gene affect carbon flux in the biosynthesis of triacylglycerols by the green alga Chlamydomonas reinhardtii[J].Biotechnology Letters,2014,36(11):2199-2208.
[33]DENG X,LI Y,FEI X.The mRNA abundance of pepc2 gene is negatively correlated with oil content in Chlamydomonas reinhardtii[J].Biomass&Bioenergy,2011,35(5):1811-1817.
[34]郑先虎,辛苑,刘航,等.一株糖蜜酒精废水净化菌Lysinibacilus sp.S6的筛选与鉴定[J].工业微生物,2017,47(3):43-48.ZHENG Xianhu,XIN Yuan,LIU Hang,et al.Screening and identification of a strain Lysinibacilus sp.S6 with purifying capability of molasses alcohol wastewater[J].Industrial Microbiology,2017,47(3):43-48.
[35]SINGH N,PETRINIC I,HéLIX-NIELSEN C,et al.Concentrating molasses distillery wastewater using biomimetic forward osmosis(FO)membranes[J].Water Research,2018,130:271-280.
[36]孙静,赵冲,武帅,等.浅议空气中二氧化碳吸收的方法[J].丝路视野,2017(6):139-140.SUN Jing,ZHAO Chong,WU Shuai,et al.Research on the absorption of carbon dioxide in the air[J].The Silk Road Vision,2017(6):139-140.
[37]ALI P,ELENA I,FELIPE R,et al.A chemical approach to manipulate the algal growth,lipid content and high-value alpha-linolenic acid for biodiesel production[J].Algal Research,2017,26:312-322.
[38]CHOKSHI K,PANCHA I,GHOSH A,et al.Microalgal biomass generation by phytoremediation of dairy industry wastewater:an integrated approach towards sustainable biofuel production[J].Bioresource Technology,2016,221:455-460.
[39]KUMAR K,MISHRA S K,SHRIVASTAV A,et al.Recent trends in the mass cultivation of algae in raceway ponds[J].Renewable and Sustainable Energy Reviews,2015,51:875-885.
[40]张芳,程丽华,徐新华,等.能源微藻采收及油脂提取技术[J].化学进展,2012,24(10):2062-2072.ZHANG Fang,CHENG Lihua,XU Xinhua,et al.Technologies of microalgal harvesting and lipid extraction[J].Progress in Chemistry,2012,24(10):2062-2072.
[2]WU X,LEUNG D Y C.Optimization of biodiesel production from camelina oil using orthogonal experiment[J].Applied Energy,2011,88(11):3615-3624.
[3]李川,胡献国.绿色生物质灰催化剂用于合成生物柴油[J].石油学报(石油加工),2018,34(1):78-85.LI Chuan,HU Xianguo.Green biomass ash catalyst for biodiesel synthesis[J].Journal of Petroleum(Petroleum Processing),2018,34(1):78-85.
[4]王常文,崔方方,宋宇.生物柴油的研究现状及发展前景[J].中国油脂,2014,39(5):44-48.WANG Changwen,CUI Fangfang,SONG Yu.Research situation and development prospect of biodiesel[J].Chinese Oils and Fats,2014,39(5):44-48.
[5]STEPHENSON P G,MOORE C M,TERRY M J,et al.Improving photosynthesis for algal biofuels:toward a green revolution[J].Trends in Biotechnology,2011,29(12):615-623.
[6]CARIOCA J O,HILUY FILHO J J,LEAL M R,et al.The hard choice for alternative biofuels to diesel in Brazil[J].Biotechnology Advances,2009,27(6):1043-1050.
[7]郭丹,银建中.微藻制备生物柴油的技术进展[J].化工装备技术,2014,35(4):4-9.GUO Dan,YIN Jianzhong.Technology progress of biodiesel production from microalgae[J].Chemical Equipment Technology,2014,35(4):4-9.
[8]方正,吕德义.微藻制备生物柴油的研究进展[J].现代化工,2017,37(9):57-61.FANG Zheng,LüDeyi.Research progress on biodiesel production by microalgae[J].Modern Chemical Industry,2017,37(9):57-61.
[9]李斐,白净,常春,等.微藻生物柴油成本控制技术进展[J].现代化工,2015,35(5):16-20.LI Fei,BAI Jing,CHANG Chun,et al.Developments of cost control technologies to produce biodiesel from microalgae[J].Modern Chemical Industry,2015,35(5):16-20.
[10]NAYAK M,KAREMORE A,SEN R.Performance evaluation of microalgae for concomitant wastewater bioremediation,CO2,biofixation and lipid biosynthesis for biodiesel application[J].Algal Research,2016,16:216-223.
[11]FERNáNDEZLINARES L C,GUERRERO B C,DURáN P E,et al.Assessment of Chlorella vulgaris and indigenous microalgae biomass with treated wastewater as growth culture medium[J].Bioresource Technology,2017,244(Pt 1):400-406.
[12]周祖光.糖蜜酒精生产废醪液资源化利用探析[J].环境科学与技术,2005,28(2):98-100.ZHOU Zuguang.Research on the utilization of molasses waste water[J].Environmental Science and Technology,2005,28(2):98-100.
[13]张跃彬,高正卿.蔗糖副产物滤泥、糖蜜酒精废醪液的开发利用[J].中国糖料,2009(1):63-64.ZHANG Yuebin,GAO Zhengqing.Development and utilization on filter mud of cane sugar by-product and wastewater from alcoholic fermentation of molasses[J].Sugar Crops of China,2009(1):63-64.
[14]CHAO M,WEN H,XING D,et al.Molasses wastewater treatment and lipid production at low temperature conditions by a microalgal mutant Scenedesmus sp.Z-4[J].Biotechnology for Biofuels,2017,10(1):111-123.
[15]YU Z,PEI H Y,JIANG L Q,et al.Phytohormone addition coupled with nitrogen depletion almost tripled the lipid productivities in two algae[J].Bioresource Technology,2018,247:904-914.
[16]ZHANG H,ZHANG N,YANG R,et al.Melatonin promotes seed germination under high salinity by regulating antioxidant systems,ABA and GA4 interaction in cucumber(Cucumis sativus L.)[J].Journal of Pineal Research,2015,57(3):269-279.
[17]BYEON Y,BACK K.An increase in melatonin in transgenic rice causes pleiotropic phenotypes,including enhanced seedling growth,delayed flowering,and low grain yield[J].Journal of Pineal Research,2014,56(4):408-414.
[18]NAWAZ M A,JIAO Y,CHEN C,et al.Melatonin pretreatment improves vanadium stress tolerance of watermelon seedlings by reducing vanadium concentration in the leaves and regulating melatonin biosynthesis and antioxidant-related gene expression[J].Journal of Plant Physiology,2018,220:115-127.
[19]YU X,ZHAO P,HE C,et al.Isolation of a novel strain of Monoraphidium sp and characterization of its potential application as biodiesel feedstock[J].Bioresource Technology,2012,121(7):256-262.
[20]CHE R,HUANG L,XU J W,et al.Effect of fulvic acid induction on the physiology,metabolism,and lipid biosynthesis-related gene transcription of Monoraphidium sp.FXY-10[J].Bioresource Technology,2016,227:324-334.
[21]BERGES J A,FISHER A E,HARRISON P J.A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana[J].Marine Biology,1993,115(2):187-193.
[22]ZHAO Y,LI D,DING K,et al.Production of biomass and lipids by the oleaginous microalgae Monoraphidium sp.QLY-1 through heterotrophic cultivation and photo-chemical modulator induction[J].Bioresour Technol,2016,211:669-676.
[23]ZHAO Y,LI D,XU J W,et al.Melatonin enhances lipid production in Monoraphidium sp.QLY-1 under nitrogen deficiency conditions via a multi-level mechanism[J].Bioresource Technology,2018,259:46-35.
[24]WEI W,LI Q T,CHU Y N,et al.Melatonin enhances plant growth and abiotic stress tolerance in soybean plants[J].Journal of Experimental Botany,2015,66(3):695-707.
[25]SHI L X,THEG S M.The chloroplast protein import system:from algae to trees[J].Biochim Biophys Acta,2013,1833(2):314-331.
[26]金永华,周伟能,吴玉林.乙酰辅酶A羧化酶抑制剂的研究进展[J].海峡药学,2018,30(1):9-14.JIN Yonghua,ZHOU Weineng,WU Yulin.Progress in the research of acetyl coenzyme a carboxylase inhibitors[J].Strait Pharmaceutical Journal,2018,30(1):9-14.
[27]GOODENOUG H U,BLABY I,CASERO D,et al.The path to triacylglyceride obesity in the sta6 strain of Chlamydomonas reinhardtii[J].Eukaryotic Cell,2014,13(5):591-613.
[28]MA X,LIU J,LIU B,et al.Physiological and biochemical changes reveal stress-associated photosynthetic carbon partitioning into triacylglycerol in the oleaginous marine alga Nannochloropsis oculata[J].Algal Research,2016,16:28-35
[29]吕娜娜,朱葆华,鹿琳,等.过表达三角褐指藻苹果酸酶基因提高E.coli脂肪酸合成能力研究[J].中国海洋大学学报(自然科学版),2016,46(5):65-69.LüNana,ZHU Baohua,LU Lin,et al.Overexpression of malic enzyme gene from phaeodactylum tricornutum promotes fatty acids production in Escherichia coli[J].Periodical of Ocean University of China(Natural Science Edition),2016,46(5):65-69.
[30]COURCHESNE N,PARISIEN A,WANG B,et al.Enhancement of lipid production using biochemical,genetic and transcription factor engineering approaches[J].Journal of Biotechnology,2009,142(1):31-41.
[31]霍垲,陆巍,李霞.干旱胁迫下调节ATP的含量对提高转玉米C4型pepc水稻光合速率的影响[J].中国生态农业学报,2015,23(5):605-613.HUO Kai,LU Wei,LI Xia.Effect of regulating ATP on improving photosynthetic rate of transgenic rice with overexpressing maize C4pepc under drought stress[J].Chinese Journal of Eco-Agriculture,2015,23(5):605-613.
[32]DENG X,CAI J,LI Y,et al.Expression and knockdown of the PEPC1gene affect carbon flux in the biosynthesis of triacylglycerols by the green alga Chlamydomonas reinhardtii[J].Biotechnology Letters,2014,36(11):2199-2208.
[33]DENG X,LI Y,FEI X.The mRNA abundance of pepc2 gene is negatively correlated with oil content in Chlamydomonas reinhardtii[J].Biomass&Bioenergy,2011,35(5):1811-1817.
[34]郑先虎,辛苑,刘航,等.一株糖蜜酒精废水净化菌Lysinibacilus sp.S6的筛选与鉴定[J].工业微生物,2017,47(3):43-48.ZHENG Xianhu,XIN Yuan,LIU Hang,et al.Screening and identification of a strain Lysinibacilus sp.S6 with purifying capability of molasses alcohol wastewater[J].Industrial Microbiology,2017,47(3):43-48.
[35]SINGH N,PETRINIC I,HéLIX-NIELSEN C,et al.Concentrating molasses distillery wastewater using biomimetic forward osmosis(FO)membranes[J].Water Research,2018,130:271-280.
[36]孙静,赵冲,武帅,等.浅议空气中二氧化碳吸收的方法[J].丝路视野,2017(6):139-140.SUN Jing,ZHAO Chong,WU Shuai,et al.Research on the absorption of carbon dioxide in the air[J].The Silk Road Vision,2017(6):139-140.
[37]ALI P,ELENA I,FELIPE R,et al.A chemical approach to manipulate the algal growth,lipid content and high-value alpha-linolenic acid for biodiesel production[J].Algal Research,2017,26:312-322.
[38]CHOKSHI K,PANCHA I,GHOSH A,et al.Microalgal biomass generation by phytoremediation of dairy industry wastewater:an integrated approach towards sustainable biofuel production[J].Bioresource Technology,2016,221:455-460.
[39]KUMAR K,MISHRA S K,SHRIVASTAV A,et al.Recent trends in the mass cultivation of algae in raceway ponds[J].Renewable and Sustainable Energy Reviews,2015,51:875-885.
[40]张芳,程丽华,徐新华,等.能源微藻采收及油脂提取技术[J].化学进展,2012,24(10):2062-2072.ZHANG Fang,CHENG Lihua,XU Xinhua,et al.Technologies of microalgal harvesting and lipid extraction[J].Progress in Chemistry,2012,24(10):2062-2072.