在明串珠菌中构建葡萄糖到甘露醇的转化体系
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摘要
为了进一步提高甘露醇产量,在明串珠菌构建了葡萄糖到甘露醇的转化体系。通过2次同源重组,将mt1d-m1p表达盒串联体定点插入到染色体上。以90 g/L蔗糖为底物时,野生型菌株的甘露醇产量为31. 48 g/L,Δaldh::(mt1d-m1p)为42. 63 g/L,Δaldh::(mt1d-m1p)Δdts::amy为43. 47 g/L,Δaldh::(mt1d-m1p)Δdts::(mt1d-m1p)为45. 74 g/L,Δdts1ΔD-ldhΔpat::mdhΔstpk::mdhΔfk::mdhΔaldh::(mt1d-m1p)为47. 26 g/L。增加甘露醇的合成途径是增产的手段之一。
In order to further increase mannitol production from sucrose in Leuconostoc sp.,a mt1d-m1p expression cassette was site-directly integrated onto its chromosome. The resulting strains,Δaldh: :(mt1d-m1p),Δaldh: :(mt1d-m1p) Δdts: : amy,Δaldh: :(mt1d-m1p) Δdts: :(mt1d-m1p),and Δdts1 ΔD-ldh Δpat: : mdh Δstpk: : mdhΔfk: : mdh Δaldh: :(mt1d-m1p) produced 42. 63,43. 47,45. 74 and 47. 26 g/L mannitol from 90 g/L sucrose. In comparison,wild type only produced 31. 48 g/L mannitol. In conclusion,increasing the synthetic route of mannitol from glucose is one of the means to increase its production.
In order to further increase mannitol production from sucrose in Leuconostoc sp.,a mt1d-m1p expression cassette was site-directly integrated onto its chromosome. The resulting strains,Δaldh: :(mt1d-m1p),Δaldh: :(mt1d-m1p) Δdts: : amy,Δaldh: :(mt1d-m1p) Δdts: :(mt1d-m1p),and Δdts1 ΔD-ldh Δpat: : mdh Δstpk: : mdhΔfk: : mdh Δaldh: :(mt1d-m1p) produced 42. 63,43. 47,45. 74 and 47. 26 g/L mannitol from 90 g/L sucrose. In comparison,wild type only produced 31. 48 g/L mannitol. In conclusion,increasing the synthetic route of mannitol from glucose is one of the means to increase its production.
引文
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[2]ZHANG M,GU L,CHENG C,et al.High-yield production of mannitol by Leuconostoc pseudomesenteroides CTCCG123 from chicory-derived inulin hydrolysate[J].Journal of Industrial Microbiology&Biotechnology,2017,44(8):1 237-1 244.
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[4]ORTIZ M E,RAYA R R,MOZZI F.Efficient mannitol production by wild-type Lactobacillus reuteri CRL 1101 is attained at constant p H using a simplified culture medium[J].Applied Microbiology and Biotechnology,2015,99(20):8 717-8 729.
[5]朱婧,吴昊,任心怡,等.碳氮源及p H调控对假肠膜明串珠菌产甘露醇的影响[J].化工进展,2015,34(12):4 333-4 337.
[6]RAKICKA M,RYWIN'SKA A,CYBULSKI K,et al.Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants[J].Brazilian Journal of Microbiology,2016,47(2):417-423.
[7]RODRGUEZ L G R,ALLER K,BRU E,et al.Enhanced mannitol biosynthesis by the fruit origin strain Fructobacillus tropaeoli CRL 2034[J].Applied Microbiology and Biotechnology,2017,101(15):6 165-6 177.
[8]MENG Q,ZHANG T,WEI W,et al.Production of mannitol from a high concentration of Glucose by Candida parapsilosis SK26.001[J].Applied Biochemistry and Biotechnology,2017,181(1):391-406.
[9]ZHANG M,GU L,CHENG C,et al.Recent advances in microbial production of mannitol:Utilization of low-cost substrates,strain development and regulation strategies[J].World Journal of Microbiology and Biotechnology,2018,34:41.
[10]PENG Y W,JIN H X.Effect of the pat,fk,stpk gene knock-out and mdh gene knock-in on mannitol production in Leuconostoc mesenteroides[J].Journal of Microbiology and Biotechnology,2018,28(12):2 009-2 018.
[11]金红星,卢哲,成文玉.一株高产甘露醇的肠膜明串珠菌突变菌株及其应用方法:中国,CN201711169481[P].2018-04-06.
[12]RESHAMWALA S M S,PAGAR S K,VELHAL V S,et al.Construction of an efficient Escherichia coli whole-cell biocatalyst for D-mannitol production[J].Journal of Bioscience and Bioengineering,2014,118(6):628-631.
[13]MAKAROVA K,SLESAREV A,WOLF Y,et al.Leuconostoc mesenteroides subsp.mesenteroides ATCC 8293,complete genome[DB].https://www.ncbi.nlm.nih.gov/nuccore/CP000414.1,[2014-01-18].
[14]田云飞,刘晓莉,金红星,等.α-淀粉酶基因在肠膜明串珠菌基因表达中的应用[J].食品工业科技,2016,37(10):203-207.
[15]ZHANG Z,CHENG W Y,JU X Y,et al.The Effect of dextransucrase gene inactivation on mannitol production by Leuconostoc mesenteroides[J].Indian Journal of Microbiology,2015,55(1):35-40.
[16]朱方剑,陈建,林丽萍,等.HPLC-ELSD法测定乌灵菌粉中甘露醇的含量[J].中国药事,2013,27(5):534-536.
[17]KOH D W,PARK J W,BLIM J,et al.A rapid method for simultaneous of 13 sugars and sugar alcohols in food products by UPLC-ELSD[J].Food Chemistry,2018,240(2):694-700.
[18]金红星,史建波,成文玉,等.以蔗糖为原料明串珠菌发酵生产甘露醇[J].食品与发酵工业,2011,37(2):91-93.
[19]金红星,成文玉,张舟.一株明串珠菌突变菌株及其构建方法和应用方法:中国,CN201410065372[P].2015-12-02.
[20]金红星,王立东,成文玉.一株产甘露醇的肠膜明串珠菌突变菌株及其应用方法:中国,CN201710051539[P].2017-05-31.
[21]鄢明辉,吴正钧.肠膜明串珠菌胞外多糖合成的蛋白组学分析[J].食品工业科技,2016,37(14):158-160;180.
[22]李海超.以红糖/甘蔗汁为原料明串珠菌发酵产甘露醇的研究[D].天津:河北工业大学,2015.
[23]罗希,曹海龙,张卉妍,等.以菊粉为底物全细胞催化生产甘露醇[J].大连工业大学学报,2017,36(4):235-239.
[24]ZHANG M,GU L,CHENG C,et al.High-yield production of mannitol by Leuconostoc pseudomesenteroides CTCCG123 from chicory-derived inulin hydrolysate[J].Journal of Industrial Microbiology&Biotechnoogy,2017,44(8):1 237-1 244.
[25]SAHA B C.Production of mannitol from inulin by simultaneous enzymatic saccharification and fermentation with Lactobacillus intermedius NRRL B-3693[J].Enzyme and Microbial Technology,2006,39(5):991-995.
[2]ZHANG M,GU L,CHENG C,et al.High-yield production of mannitol by Leuconostoc pseudomesenteroides CTCCG123 from chicory-derived inulin hydrolysate[J].Journal of Industrial Microbiology&Biotechnology,2017,44(8):1 237-1 244.
[3]OTGONBAYAR G E,EOM H J,KIM B S,et al.Mannitol production by Leuconostoc citreum KACC 91348P Isolated from Kimchi[J].Journal of Microbiology and Biotechnology,2011,21(9):968-971.
[4]ORTIZ M E,RAYA R R,MOZZI F.Efficient mannitol production by wild-type Lactobacillus reuteri CRL 1101 is attained at constant p H using a simplified culture medium[J].Applied Microbiology and Biotechnology,2015,99(20):8 717-8 729.
[5]朱婧,吴昊,任心怡,等.碳氮源及p H调控对假肠膜明串珠菌产甘露醇的影响[J].化工进展,2015,34(12):4 333-4 337.
[6]RAKICKA M,RYWIN'SKA A,CYBULSKI K,et al.Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants[J].Brazilian Journal of Microbiology,2016,47(2):417-423.
[7]RODRGUEZ L G R,ALLER K,BRU E,et al.Enhanced mannitol biosynthesis by the fruit origin strain Fructobacillus tropaeoli CRL 2034[J].Applied Microbiology and Biotechnology,2017,101(15):6 165-6 177.
[8]MENG Q,ZHANG T,WEI W,et al.Production of mannitol from a high concentration of Glucose by Candida parapsilosis SK26.001[J].Applied Biochemistry and Biotechnology,2017,181(1):391-406.
[9]ZHANG M,GU L,CHENG C,et al.Recent advances in microbial production of mannitol:Utilization of low-cost substrates,strain development and regulation strategies[J].World Journal of Microbiology and Biotechnology,2018,34:41.
[10]PENG Y W,JIN H X.Effect of the pat,fk,stpk gene knock-out and mdh gene knock-in on mannitol production in Leuconostoc mesenteroides[J].Journal of Microbiology and Biotechnology,2018,28(12):2 009-2 018.
[11]金红星,卢哲,成文玉.一株高产甘露醇的肠膜明串珠菌突变菌株及其应用方法:中国,CN201711169481[P].2018-04-06.
[12]RESHAMWALA S M S,PAGAR S K,VELHAL V S,et al.Construction of an efficient Escherichia coli whole-cell biocatalyst for D-mannitol production[J].Journal of Bioscience and Bioengineering,2014,118(6):628-631.
[13]MAKAROVA K,SLESAREV A,WOLF Y,et al.Leuconostoc mesenteroides subsp.mesenteroides ATCC 8293,complete genome[DB].https://www.ncbi.nlm.nih.gov/nuccore/CP000414.1,[2014-01-18].
[14]田云飞,刘晓莉,金红星,等.α-淀粉酶基因在肠膜明串珠菌基因表达中的应用[J].食品工业科技,2016,37(10):203-207.
[15]ZHANG Z,CHENG W Y,JU X Y,et al.The Effect of dextransucrase gene inactivation on mannitol production by Leuconostoc mesenteroides[J].Indian Journal of Microbiology,2015,55(1):35-40.
[16]朱方剑,陈建,林丽萍,等.HPLC-ELSD法测定乌灵菌粉中甘露醇的含量[J].中国药事,2013,27(5):534-536.
[17]KOH D W,PARK J W,BLIM J,et al.A rapid method for simultaneous of 13 sugars and sugar alcohols in food products by UPLC-ELSD[J].Food Chemistry,2018,240(2):694-700.
[18]金红星,史建波,成文玉,等.以蔗糖为原料明串珠菌发酵生产甘露醇[J].食品与发酵工业,2011,37(2):91-93.
[19]金红星,成文玉,张舟.一株明串珠菌突变菌株及其构建方法和应用方法:中国,CN201410065372[P].2015-12-02.
[20]金红星,王立东,成文玉.一株产甘露醇的肠膜明串珠菌突变菌株及其应用方法:中国,CN201710051539[P].2017-05-31.
[21]鄢明辉,吴正钧.肠膜明串珠菌胞外多糖合成的蛋白组学分析[J].食品工业科技,2016,37(14):158-160;180.
[22]李海超.以红糖/甘蔗汁为原料明串珠菌发酵产甘露醇的研究[D].天津:河北工业大学,2015.
[23]罗希,曹海龙,张卉妍,等.以菊粉为底物全细胞催化生产甘露醇[J].大连工业大学学报,2017,36(4):235-239.
[24]ZHANG M,GU L,CHENG C,et al.High-yield production of mannitol by Leuconostoc pseudomesenteroides CTCCG123 from chicory-derived inulin hydrolysate[J].Journal of Industrial Microbiology&Biotechnoogy,2017,44(8):1 237-1 244.
[25]SAHA B C.Production of mannitol from inulin by simultaneous enzymatic saccharification and fermentation with Lactobacillus intermedius NRRL B-3693[J].Enzyme and Microbial Technology,2006,39(5):991-995.