齐整小核菌SCL2010产小核菌多糖培养基优化的研究
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摘要
采用单因素和正交试验对小核菌多糖高产菌株齐整小核菌(Sclerotium rolfsii)SCL2010的培养基成分进行了深入地筛选与优化。结果表明,最佳培养基配方为葡萄糖45.0 g/L,玉米浆1.5 g/L,NaNO_32.0 g/L,K_2HPO_4·3H_2O 2.0 g/L,柠檬酸0.7 g/L,MgSO_4·7H_2O1.5 g/L,KCl 2.0 g/L。在此优化条件下,小核菌多糖的产量为31.81 g/L,碳源转化率为70.69%。采用发酵罐进行小试放大试验,小核菌多糖的产量达到31.86 g/L,碳源转化率为70.80%,发酵液表观黏度达到4 500 m Pa·s,并将发酵时间缩短至60 h左右,具有显著效果。
The medium components of high scleroglucan-producing Sclerotium rolfsii SCL2010 were deeply screened and optimized by single factor and orthogonal experiments. The results showed that the optimum medium formula was determined as follows: glucose 45.0 g/L, corn steep liquor1.5 g/L, NaNO_3 2.0 g/L, K_2HPO_4·3H_2O 2.0 g/L, citric acid 0.7 g/L, MgSO_4·7H_2O 1.5 g/L, KCl 2.0 g/L. Under the optimal conditions, the yield of scleroglucan was 31.81 g/L, and the conversion rate of carbon source was 70.69%. Finally, the scale-up experiment was carried out by the fermentor,the yield of scleroglucan was 31.86 g/L and the conversion rate of carbon source was 70.80%, the apparent viscosity of fermentation broth reached4 500 m Pa·s, and the fermentation time was shortened to about 60 h, which had significant breakthrough.
The medium components of high scleroglucan-producing Sclerotium rolfsii SCL2010 were deeply screened and optimized by single factor and orthogonal experiments. The results showed that the optimum medium formula was determined as follows: glucose 45.0 g/L, corn steep liquor1.5 g/L, NaNO_3 2.0 g/L, K_2HPO_4·3H_2O 2.0 g/L, citric acid 0.7 g/L, MgSO_4·7H_2O 1.5 g/L, KCl 2.0 g/L. Under the optimal conditions, the yield of scleroglucan was 31.81 g/L, and the conversion rate of carbon source was 70.69%. Finally, the scale-up experiment was carried out by the fermentor,the yield of scleroglucan was 31.86 g/L and the conversion rate of carbon source was 70.80%, the apparent viscosity of fermentation broth reached4 500 m Pa·s, and the fermentation time was shortened to about 60 h, which had significant breakthrough.
引文
[1]FARI譙A J I,SI譙ERIZ F,MOLINA O E,et al.Isolation and physicochemical characterization of soluble scleroglucan from Sclerotium rolfsii.Rheological properties,molecular weight and conformational characteristics[J].Carbohyd Polym,2001,44(1):41-50.
[2]FARI譙A J I,VI譙ARTA S C,CATTANEO M,et al.Structural stability of Sclerotium rolfsii ATCC 201126β-glucan with fermentation time:a chemical,infrared spectroscopic and enzymatic approach[J].J Appl Microbiol,2009,106(1):221-232.
[3]VI譙ARTA S C,DELGADO O D,FIGUEROA L I C,et al.Effects of thermal,alkaline and ultrasonic treatments on scleroglucan stability and flow behavior[J].Carbohyd Polym,2013,94(1):496-504.
[4]SCHMID J,MEYER V,SIEBER V.Scleroglucan:biosynthesis,production and application of a versatile hydrocolloid[J].Appl Microbiol Biotechnol,2011,91(4):937-947.
[5]天津市工业微生物研究所.小核菌硬葡聚糖发酵液作为油田钻井液处理剂的应用,CN201210587334.4[P].2013-05-08.
[6]李冰,张建法,蒋鹏举.真菌硬葡聚糖的生产及在油田上的应用[J].微生物学通报,2003,30(5):99-102.
[7]杜冠乐,段凡.天然高分子在油田钻井液的应用研究[J].石油化工应用,2013,32(8):1-4.
[8]HALLIBURTON ENERGY SERVICES,INC.Inhibiting salting out of diutan or scleroglucan in well treatment:US,20140378353 A1[P].2014-12-25.
[9]天津市工业微生物研究所.小核菌硬葡聚糖在线发酵提取方法及系统,CN201110006769.0[P].2013-06-05.
[10]TASKIN M,ERDAL S,CANLI O.Utilization of waste loquat(Eriobotrya Japonica Lindley)kernels as substrate for scleroglucan production by locally isolated Sclerotium rolfsii[J].Food Sci Biotechnol,2010,19(4):1069-1075.
[11]SURVASE S A,SAUDAGAR P S,SINGHAL R S.Production of scleroglucan from Sclerotium rolfsii MTCC 2156[J].Bioresour Technol,2006,97(8):989-993.
[12]WANG Y,MCNEIL B.Scleroglucan and oxalic acid formation by Sclerotium glucanicum in sucrose supplemented fermentations[J].Biotechnol Lett,1994,16(6):605-610.
[13]FARI譙A J I,SANTOS V E,PEROTTI N I,et al.Influence of the nitrogen source on the production and rheological properties of scleroglucan produced by Sclerotium rolfsii ATCC 201126[J].World J Microbiol Biotechnol,1999,15(2):309-316.
[14]SCHILLING B M,RAU U,MAIER T,et al.Modeling and scale-up of the unsterile scleroglucan production process with Sclerotium rolfsii ATCC 15205[J].Bioprocess Eng,1999,20(3):195-201.
[15]SURVASE S A,SAUDAGAR P S,SINGHAL R S.Use of complex media for the production of scleroglucan by Sclerotium rolfsii MTCC 2156[J].Bioresource Technol,2007,98(7):1509-1512.
[16]FARI譙A J I,SI譙ERIZ F,MOLINA O E,et al.High scleroglucan production by Sclerotium rolfsii:influence of medium composition[J].Biotechnol Lett,1998,20(9):825-831.
[17]SCHILLING B M.Sclerotium rolfsii ATCC 15205 in continuous culture:Economical aspects of scleroglucan production[J].Bioprocess Eng,2000,22(1):57-61.
[18]杨林娥,彭晓光,杨庆文,等.斐林试剂法测定还原糖方法的改进[J].中国酿造,2010,29(5):160-161.
[2]FARI譙A J I,VI譙ARTA S C,CATTANEO M,et al.Structural stability of Sclerotium rolfsii ATCC 201126β-glucan with fermentation time:a chemical,infrared spectroscopic and enzymatic approach[J].J Appl Microbiol,2009,106(1):221-232.
[3]VI譙ARTA S C,DELGADO O D,FIGUEROA L I C,et al.Effects of thermal,alkaline and ultrasonic treatments on scleroglucan stability and flow behavior[J].Carbohyd Polym,2013,94(1):496-504.
[4]SCHMID J,MEYER V,SIEBER V.Scleroglucan:biosynthesis,production and application of a versatile hydrocolloid[J].Appl Microbiol Biotechnol,2011,91(4):937-947.
[5]天津市工业微生物研究所.小核菌硬葡聚糖发酵液作为油田钻井液处理剂的应用,CN201210587334.4[P].2013-05-08.
[6]李冰,张建法,蒋鹏举.真菌硬葡聚糖的生产及在油田上的应用[J].微生物学通报,2003,30(5):99-102.
[7]杜冠乐,段凡.天然高分子在油田钻井液的应用研究[J].石油化工应用,2013,32(8):1-4.
[8]HALLIBURTON ENERGY SERVICES,INC.Inhibiting salting out of diutan or scleroglucan in well treatment:US,20140378353 A1[P].2014-12-25.
[9]天津市工业微生物研究所.小核菌硬葡聚糖在线发酵提取方法及系统,CN201110006769.0[P].2013-06-05.
[10]TASKIN M,ERDAL S,CANLI O.Utilization of waste loquat(Eriobotrya Japonica Lindley)kernels as substrate for scleroglucan production by locally isolated Sclerotium rolfsii[J].Food Sci Biotechnol,2010,19(4):1069-1075.
[11]SURVASE S A,SAUDAGAR P S,SINGHAL R S.Production of scleroglucan from Sclerotium rolfsii MTCC 2156[J].Bioresour Technol,2006,97(8):989-993.
[12]WANG Y,MCNEIL B.Scleroglucan and oxalic acid formation by Sclerotium glucanicum in sucrose supplemented fermentations[J].Biotechnol Lett,1994,16(6):605-610.
[13]FARI譙A J I,SANTOS V E,PEROTTI N I,et al.Influence of the nitrogen source on the production and rheological properties of scleroglucan produced by Sclerotium rolfsii ATCC 201126[J].World J Microbiol Biotechnol,1999,15(2):309-316.
[14]SCHILLING B M,RAU U,MAIER T,et al.Modeling and scale-up of the unsterile scleroglucan production process with Sclerotium rolfsii ATCC 15205[J].Bioprocess Eng,1999,20(3):195-201.
[15]SURVASE S A,SAUDAGAR P S,SINGHAL R S.Use of complex media for the production of scleroglucan by Sclerotium rolfsii MTCC 2156[J].Bioresource Technol,2007,98(7):1509-1512.
[16]FARI譙A J I,SI譙ERIZ F,MOLINA O E,et al.High scleroglucan production by Sclerotium rolfsii:influence of medium composition[J].Biotechnol Lett,1998,20(9):825-831.
[17]SCHILLING B M.Sclerotium rolfsii ATCC 15205 in continuous culture:Economical aspects of scleroglucan production[J].Bioprocess Eng,2000,22(1):57-61.
[18]杨林娥,彭晓光,杨庆文,等.斐林试剂法测定还原糖方法的改进[J].中国酿造,2010,29(5):160-161.