玉米淀粉浓度对酶法制备直链麦芽低聚糖的影响
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摘要
为研究浓度对玉米淀粉酶解过程和产物的影响,以直链麦芽低聚糖含量和产率为指标,探究了直链麦芽低聚糖生成酶(Bst-MFA酶)对不同浓度(质量分数)玉米淀粉的酶解过程,并对相关机理进行了分析。结果表明,玉米淀粉质量分数由10%提高至50%时,酶解24 h后直链麦芽低聚糖和主产物麦芽五糖、麦芽六糖的含量均呈现逐渐升高的趋势,且二者均在45%的底物质量分数时达到最大值,分别为344. 47和192. 33 mg/g;而直链麦芽低聚糖的产率逐渐降低。其主要原因可能是:随底物浓度的增加,液化和糖化过程中体系黏度增大,水分子运动性降低,体系中自由水明显减少,且酶在不同浓度玉米淀粉中的作用模式存在差异,使得高浓度条件下其对玉米淀粉的水解作用不够充分,直链麦芽低聚糖的产率呈下降趋势。该研究可为工业生产中直链麦芽低聚糖的酶法制备提供参考依据。
Linear malto-oligosaccharides can be produced from corn starches by malto-oligosaccharides-forming amylase( Bst-MFAse). This study aimed to evaluate the effects of initial substrate concentration on producing linear malto-oligosaccharides by Bst-MFAse using the contents and yields of linear malto-oligosaccharides as indicators,followed by analyzing related mechanisms. The results showed that when the initial concentration of corn starch increased from 10% to 50%( w/w),the contents of linear malto-oligosaccharides and main products( maltopentaose and maltohexaose) increased gradually after hydrolyzing for 24 h. Besides,their contents reached the highest with 45% substrate,which were 344. 47 and 192. 33 mg/g,respectively. However,the yield of linear malto-oligosaccharides decreased gradually. This might be due to the increased substrate concentration enhanced the viscosity of the system during liquefaction and saccharification,which resulted in decreased water molecule mobility and significantly reduced free water amount. Additionally,MFAse had different action modes among different concentrations of corn starches,which could cause insufficient hydrolysis of corn starch molecules at high concentration. Overall,this study provides a reference for enzymatic preparation of linear malto-oligosaccharides in industries.
Linear malto-oligosaccharides can be produced from corn starches by malto-oligosaccharides-forming amylase( Bst-MFAse). This study aimed to evaluate the effects of initial substrate concentration on producing linear malto-oligosaccharides by Bst-MFAse using the contents and yields of linear malto-oligosaccharides as indicators,followed by analyzing related mechanisms. The results showed that when the initial concentration of corn starch increased from 10% to 50%( w/w),the contents of linear malto-oligosaccharides and main products( maltopentaose and maltohexaose) increased gradually after hydrolyzing for 24 h. Besides,their contents reached the highest with 45% substrate,which were 344. 47 and 192. 33 mg/g,respectively. However,the yield of linear malto-oligosaccharides decreased gradually. This might be due to the increased substrate concentration enhanced the viscosity of the system during liquefaction and saccharification,which resulted in decreased water molecule mobility and significantly reduced free water amount. Additionally,MFAse had different action modes among different concentrations of corn starches,which could cause insufficient hydrolysis of corn starch molecules at high concentration. Overall,this study provides a reference for enzymatic preparation of linear malto-oligosaccharides in industries.
引文
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[17] BIJTTEBIER A,GOESAERT H,DELCOUR J A. Temperature impacts the multiple attack action of amylases[J].Biomacromolecules,2007,8(3):765-772.
[18] CHENG Shasha,ZHANG Tan,YAO Li,et al. Use of low field-NMR and MRI to characterize water mobility and distribution in Pacific oyster(Crassostrea gigas)during drying process[J]. Drying Technology,2017,36(5):630-636.
[19] XU Danping,HONG Yan,GU Zhengbiao,et al. Effect of high pressure steam on the eating quality of cooked rice[J]. LWT-Food Science and Technology,2019,104:100-108.
[20] LI Chun,ZHANG Luda,REN Fazheng,et al. Study on different chilling factors influencing water-holding capacity of pork based on low-field nuclear magnetic resonance(LF-NMR)[J]. Transactions of the Chinese Society of Agricultural Engineering,2012,28(23):243-249.
[21]闫晓蕾.山梨糖醇在香肠制品中的持水作用研究[D].无锡:江南大学,2012.
[22]张莉莉,张涛,姜晓明,等.水分状态对鱼糜凝胶高温处理热稳定性的影响[J].食品与发酵工业,2013,39(2):56-61.
[23]赵骏倢.高效液化糖化高浓度玉米淀粉关键酶系的优化[D].长春:吉林大学,2015.
[24] GOESAERT H,BIJTTEBIER A,DELCOUR J A. Hydrolysis of amylopectin by amylolytic enzymes:Level of inner chain attack as an important analytical differentiation criterion[J]. Carbohydrate Research,2010,345(3):397-401.
[25] BAILEY J M,WHELAN W J. Physical properties of starch. I. Relationship between iodine stain and chain length[J]. Journal of Biological Chemistry,1961,236(4):969-973.
[26] KONG Haocun,ZOU Yingying,GU Zhengbiao,et al. Liquefaction concentration impacts the fine structure of maltodextrin[J]. Industrial Crops and Products,2018,123:687-697.
[2] ALI M B,MHIRI S,MEZGHANI M,et al. Purification and sequence analysis of the atypical maltohexaose-formingα-amylase of the B. stearothermophilus US100[J]. Enzyme&Microbial Technology,2001,28(6):537-542.
[3]毛跟年,杨亚洲,许牡丹,等.磷酸寡糖前体———麦芽低聚糖制备工艺的研究[J].陕西科技大学学报:自然科学版,2003,21(6):21-24.
[4] PANG Xin,XIAO Qicai,CHENG Yi,et al. Bacteria-responsive nanoliposomes as smart sonotheranostics for multidrug resistant bacterial infections[J]. ACS Nano,2019,13(2):2 427-2 438.
[5] YUN J W,LEE M G,SONG S K. Continuous production of isomalto-oligosaccharides from maltose syrup by immobilized cells of permeabilized Aureobasidium pullulans[J].Biotechnology Letters,1994,16(11):1 145-1 150.
[6] MARCHAL L M,TRAMPER J. Hydrolytic gain during hydrolysis reactions; implications and correction procedures[J]. Biotechnology Techniques,1999,13(5):325-328.
[7] QUAX W J,MRABET N T,LUITEN R G,et al. Enhancing the thermostability of glucose isomerase by protein engineering[J]. Bio/technology,1991,9(8):738-742.
[8]彭丹丹,复合酶液化高浓度玉米淀粉乳的研究[D].无锡:江南大学,2015.
[9]杨倩雯.高浓度底物条件下酶法生产麦芽糖浆工艺的研究[D].无锡:江南大学,2017.
[10]曲世洋,李才明,顾正彪,等.底物浓度对糖化反应过程的影响[J].食品与发酵工业,2014,40(3):8-12.
[11] PAN Sihui,DING Ning,REN Junyan,et al. Maltooligosaccharide-forming amylase:Characteristics,preparation,and application[J]. Biotechnology Advances,2017,35(5):619-632.
[12]潘思惠.直链麦芽低聚糖生成酶在枯草芽孢杆菌中的分泌表达及其酶学性质与产物合成研究[D].无锡:江南大学,2018.
[13] MILLER G L. Use of dinitrosalicylic acid reagent for determination of reducing sugar[J]. Analytical Biochemistry,1959,31(3):426-428.
[14]刘文静,李兆丰,顾正彪,等.微波预处理对玉米淀粉液化的影响[J].食品与发酵工业,2013,39(1):21-25.
[15] ROUSSEL X,LANCELON-PIN C,VIKS-NIELSEN A,et al. Characterization of substrate and product specificity of the purified recombinant glycogen branching enzyme of Rhodothermus obamensis[J]. Biochimica et Biophysica Acta(BBA)-General Subjects,2013,1 830(1):2 167-2 177.
[16]刘文静.酶法液化高浓度玉米淀粉乳的研究[D].无锡:江南大学,2013.
[17] BIJTTEBIER A,GOESAERT H,DELCOUR J A. Temperature impacts the multiple attack action of amylases[J].Biomacromolecules,2007,8(3):765-772.
[18] CHENG Shasha,ZHANG Tan,YAO Li,et al. Use of low field-NMR and MRI to characterize water mobility and distribution in Pacific oyster(Crassostrea gigas)during drying process[J]. Drying Technology,2017,36(5):630-636.
[19] XU Danping,HONG Yan,GU Zhengbiao,et al. Effect of high pressure steam on the eating quality of cooked rice[J]. LWT-Food Science and Technology,2019,104:100-108.
[20] LI Chun,ZHANG Luda,REN Fazheng,et al. Study on different chilling factors influencing water-holding capacity of pork based on low-field nuclear magnetic resonance(LF-NMR)[J]. Transactions of the Chinese Society of Agricultural Engineering,2012,28(23):243-249.
[21]闫晓蕾.山梨糖醇在香肠制品中的持水作用研究[D].无锡:江南大学,2012.
[22]张莉莉,张涛,姜晓明,等.水分状态对鱼糜凝胶高温处理热稳定性的影响[J].食品与发酵工业,2013,39(2):56-61.
[23]赵骏倢.高效液化糖化高浓度玉米淀粉关键酶系的优化[D].长春:吉林大学,2015.
[24] GOESAERT H,BIJTTEBIER A,DELCOUR J A. Hydrolysis of amylopectin by amylolytic enzymes:Level of inner chain attack as an important analytical differentiation criterion[J]. Carbohydrate Research,2010,345(3):397-401.
[25] BAILEY J M,WHELAN W J. Physical properties of starch. I. Relationship between iodine stain and chain length[J]. Journal of Biological Chemistry,1961,236(4):969-973.
[26] KONG Haocun,ZOU Yingying,GU Zhengbiao,et al. Liquefaction concentration impacts the fine structure of maltodextrin[J]. Industrial Crops and Products,2018,123:687-697.