黑曲霉H9-30全细胞催化合成低聚异麦芽糖
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摘要
以黑曲霉H9-30全细胞为催化剂转化麦芽糖生产低聚异麦芽糖,通过单因素实验确定其摇瓶最佳转化条件为:温度48℃,初始p H值4. 2,麦芽糖质量浓度为600 g/L,黑曲霉细胞添加量为15 g/L。此条件下,有效三糖(异麦芽糖、潘糖和异麦芽三糖)占总糖质量分数的50. 5%,总低聚异麦芽糖含量为63. 3%,达到低聚异麦芽糖工业生产标准。研究结果表明,利用黑曲霉全细胞催化生产低聚异麦芽糖具有较好的操作稳定性,生产IMO-50的半衰期达到20批次(10 d),有望应用于工业化生产低聚异麦芽糖。
Isomaltooligosaccharides( IMOs) are a kind of functional oligosaccharides that have been widely used in food industries. A new procedure for IMOs preparation was developed by using whole-cell of Aspergillus niger H9-30 as a catalyst and maltose as substrate. The optimal reaction condition in shaking flasks was as follows: at 48 ℃with an initial pH of 4. 2,with 600 g/L maltose and 15 g/L A. niger H9-30. Under this condition,the yields of isomaltose,panose,and isomaltotriose accounted for 50. 5% of total sugar,and total IMOs content accounted for63. 3%. It was also found that using whole-cell of A. niger to produce IMOs had good operational stability with the half-life reached 20 batches( 10 d). In conclusion,A. niger H9-30 has potential use for industrial production of IMO.
Isomaltooligosaccharides( IMOs) are a kind of functional oligosaccharides that have been widely used in food industries. A new procedure for IMOs preparation was developed by using whole-cell of Aspergillus niger H9-30 as a catalyst and maltose as substrate. The optimal reaction condition in shaking flasks was as follows: at 48 ℃with an initial pH of 4. 2,with 600 g/L maltose and 15 g/L A. niger H9-30. Under this condition,the yields of isomaltose,panose,and isomaltotriose accounted for 50. 5% of total sugar,and total IMOs content accounted for63. 3%. It was also found that using whole-cell of A. niger to produce IMOs had good operational stability with the half-life reached 20 batches( 10 d). In conclusion,A. niger H9-30 has potential use for industrial production of IMO.
引文
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[22] CHEN G G,LI W,ZHANG Y K,et al. A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar[J]. World Journal of Microbiology&Biotechnology,2011,27(6):1 519-1 523.
[23]岳振峰,彭志英.固定化酶法生产低聚异麦芽糖的研究[J].食品与发酵工业,2002,28(3):6-9.
[24] VETERE A,GAMINI A,CAMPA C,et al. Regiospecific transglycolytic synthesis and structural characterization of6-O-α-glucopyranosyl-glucopyranose(Isomaltose)[J].Biochemical and Biophysical Research Communications,2000,274(1):99-104.
[25] KATO N,SUYAMA S,SHIROKANE M,et al. Novel alpha-glucosidase from Aspergillus nidulans with strong transglycosylation activity[J]. Applied&Environmental Microbiology,2002,68(3):1 250-1 256.
[26] MACKENZIE L F,WANG Q,WARREN R A J,et al.Glycosynthases:Mutant glycosidases for oligosaccharide synthesis[J]. Journal of the American Chemical Society,1998,120(22):5 583-5 584.
[2] CHEN R. Enzyme and microbial technology for synthesis of bioactive oligosaccharides:An update[J]. Applied Microbiology&Biotechnology,2018,102(7):3 017-3 026.
[3] YAN Q J,HAN P,YANG S Q,et al. Purification and characterization of a novelα-glucosidase from Malbranchea cinnamomea[J]. Biotechnology Letters,2015,37(6):1 279-1 286.
[4] MADSEN L R,STANLEY S,SWANN P,et al. A survey of commercially available isomaltooligosaccharide-based food ingredients[J]. Journal of Food Science,2017,82(2):401-408.
[5]陈桂光,李玮,齐辉连,等.黑曲霉低聚异麦芽糖高产菌株的诱变选育[J].食品工业科技,2011(8):185-187.
[6] SORNDECH W,SAGNELLI D,BLENNOW A,et al.Combination of amylase and transferase catalysis to improve IMO compositions and productivity[J]. LWT-Food Science and Technology,2017,79:479-486.
[7] RUDEEKULTHAMRONG P,SAWASDEE K,KAULPIBOON J,et al. Production of long-chain isomaltooligosaccharides from maltotriose using the thermostable amylomaltase and transglucosidase enzymes[J]. Biotechnology&Bioprocess Engineering,2013,18(4):778-786.
[8] PAN Y C,LEE W C. Production of high-purity isomaltooligosaccharides syrup by the enzymatic conversion of transglucosidase and fermentation of yeast cells[J]. Biotechnology&Bioengineering,2010,89(7):797-804.
[9]吴孔阳,齐宗献,黄桂华,等.一株产α-葡萄糖苷酶菌株的鉴定和选育[J].食品研究与开发,2012,33(12):145-149.
[10] CHEN G G,LI W,ZHANG Y K,et al. A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar[J]. World Journal of Microbiology&Biotechnology,2011,27(6):1 519-1 523.
[11] MA M,OKUYAMA M,SATO M,et al. Effects of mutation of Asn694 in Aspergillus nigerα-glucosidase on hydrolysis and transglucosylation[J]. Applied Microbiology&Biotechnology,2017,101(16):6 399-6 408.
[12] KOBAYASHI I,TOKUDA M H,HASHIMOTO H,et al.Purification and characterization of a new type of alphaglucosidase from Paecilomyces lilacinus that has transglucosylation activity to produce alpha-1,3-and alpha-1,2-linked oligosaccharides[J]. Bioscience Biotechnology&Biochemistry,2003,67(1):29-35.
[13] YAMASAKI Y,SUZUKI Y,OZAWA J. Three forms ofα-glucosidase and a glucoamylase from Aspergillus awamori[J]. Journal of the Agricultural Chemical Society of Japan,1977,41(11):2 149-2 161.
[14] ZHANG L,SU Y L,ZHENG Y,et al. Sandwich-structured enzyme membrane reactor for efficient conversion of maltose into isomaltooligosaccharides[J]. Bioresource Technology,2010,101(23):9 144-9 149.
[15] HUANG Z,LI Z,SU Y,et al. Continuous production of isomalto-oligosaccharides by thermo-inactivated cells of Aspergillus niger J2 with coarse perlite as an immobilizing material[J]. Applied Biochemistry and Biotechnology,2018,185(4):1 088-1 099.
[16] BASU A,MUTTURI S,PRAPULLA S G. Production of isomaltooligosaccharides(IMO)using simultaneous saccharification and transglucosylation from starch and sustainable sources[J]. Process Biochemistry,2016,51(10):1 464-1 471.
[17]周康,刘冬梅,范梦珂,等. Aspergillus niger CGMCC No. 6640全细胞生物转化制备蔗果低聚糖[J].现代食品科技,2013,29(8):1 967-1 972.
[18]章爱媛.黑曲霉RAF106全细胞生物转化茶多酚的研究[D].广州:华南农业大学,2016.
[19]刘国生,邢善涛,王建琨,等.埃希菌全细胞转化阿糖尿苷合成阿糖腺苷[J].中国医药工业杂志,2010,41(6):416-419.
[20] OJHA S,MISHRA S,CHAND S. Production of isomaltooligosaccharides by cell boundα-glucosidase of Microbacterium sp.[J]. LWT-Food Science and Technology,2015,60(1):486-494.
[21]徐燕杉.黑曲霉α-葡萄糖苷酶在毕赤酵母的表面展示及催化合成低聚异麦芽糖的研究[D].广州:华南理工大学,2017.
[22] CHEN G G,LI W,ZHANG Y K,et al. A high-throughput method for screening of Aspergillus niger mutants with high transglycosylation activity by detecting non-fermentable reducing sugar[J]. World Journal of Microbiology&Biotechnology,2011,27(6):1 519-1 523.
[23]岳振峰,彭志英.固定化酶法生产低聚异麦芽糖的研究[J].食品与发酵工业,2002,28(3):6-9.
[24] VETERE A,GAMINI A,CAMPA C,et al. Regiospecific transglycolytic synthesis and structural characterization of6-O-α-glucopyranosyl-glucopyranose(Isomaltose)[J].Biochemical and Biophysical Research Communications,2000,274(1):99-104.
[25] KATO N,SUYAMA S,SHIROKANE M,et al. Novel alpha-glucosidase from Aspergillus nidulans with strong transglycosylation activity[J]. Applied&Environmental Microbiology,2002,68(3):1 250-1 256.
[26] MACKENZIE L F,WANG Q,WARREN R A J,et al.Glycosynthases:Mutant glycosidases for oligosaccharide synthesis[J]. Journal of the American Chemical Society,1998,120(22):5 583-5 584.