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      Effects of Temperature and Additives on the Thermal Stability of Glucoamylase from Aspergillus niger = Effects of Temperature and Additives on the Thermal Stability of Glucoamylase from Aspergillus niger

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      https://www.riss.kr/link?id=A100284080

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      다국어 초록 (Multilingual Abstract)

      GAM-1 and GAM-2, two themostable glucoamylases from Aspergillus niger B-30, possess different molecular masses, glycosylation, and thermal stability. In the present study, the effects of additives on the thermal inactivation of GAM-1 and GAM-2 were investigated. The half-lives of GAM-1 and GAM-2 at 70ºC were 45 and 216 min, respectively. Data obtained from fluorescence spectroscopy, circular dichroism spectroscopy, UV absorption spectroscopy, and dynamic light scattering demonstrated that during the thermal inactivation progress, combined with the loss of the helical structure and a majority of the tertiary structure, tryptophan residues were partially exposed and further led to glucoamylases aggregating. The thermal stability of GAM-1 and GAM-2 was largely improved in the presence of sorbitol and trehalose. Results from spectroscopy and Native-PAGE confirmed that sorbitol and trehalose maintained the native state of glucoamylases and prevented their thermal aggregation. The loss of hydrophobic bonding and helical structure was responsible for the decrease of glucoamylase activity. Additionally, sorbitol and trehalose significantly increased the substrate affinity and catalytic efficiency of the two glucoamylases. Our results display an insight into the thermal inactivation of glucoamylases and provide an important base for industrial applications of the thermally stable glucoamylases.
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      GAM-1 and GAM-2, two themostable glucoamylases from Aspergillus niger B-30, possess different molecular masses, glycosylation, and thermal stability. In the present study, the effects of additives on the thermal inactivation of GAM-1 and GAM-2 were in...

      GAM-1 and GAM-2, two themostable glucoamylases from Aspergillus niger B-30, possess different molecular masses, glycosylation, and thermal stability. In the present study, the effects of additives on the thermal inactivation of GAM-1 and GAM-2 were investigated. The half-lives of GAM-1 and GAM-2 at 70ºC were 45 and 216 min, respectively. Data obtained from fluorescence spectroscopy, circular dichroism spectroscopy, UV absorption spectroscopy, and dynamic light scattering demonstrated that during the thermal inactivation progress, combined with the loss of the helical structure and a majority of the tertiary structure, tryptophan residues were partially exposed and further led to glucoamylases aggregating. The thermal stability of GAM-1 and GAM-2 was largely improved in the presence of sorbitol and trehalose. Results from spectroscopy and Native-PAGE confirmed that sorbitol and trehalose maintained the native state of glucoamylases and prevented their thermal aggregation. The loss of hydrophobic bonding and helical structure was responsible for the decrease of glucoamylase activity. Additionally, sorbitol and trehalose significantly increased the substrate affinity and catalytic efficiency of the two glucoamylases. Our results display an insight into the thermal inactivation of glucoamylases and provide an important base for industrial applications of the thermally stable glucoamylases.

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      참고문헌 (Reference)

      1 Miller GL, "Use of dinitrosalicylic acid reagent for determination of reducing sugar" 31 : 426-428, 1959

      2 Cui L, "Thermal stability and conformational changes of transglutaminase from a newly isolated Streptomyces hygroscopicus" 99 : 3794-3800, 2008

      3 Ca ves MS, "Thermal inactivation of uricase (urate oxidase): mechanism and effects of additives" 52 : 497-507, 2013

      4 Gangadhara, "The stabilizing effects of polyols and sugars on porcine pancreatic lipase" 86 : 773-781, 2009

      5 Negi S, "Study of conformational changes in glucoamylase of Aspergillus awamori nakazawa in presence of denaturants through CD-spectroscopy" 101 : 7577-7580, 2010

      6 Zaroog MS, "Stabilizing effect of various polyols on the native and the denatured states of glucoamylase" 2013

      7 Chi MC, "Sorbitol counteracts temperature and chemical-induced denaturation of a recombinant α-amylase from alkaliphilic Bacillus sp. TS-23" 39 : 1779-1788, 2012

      8 D ong XY, "Self-interaction of native and denatured lysozyme in the presence of osmolytes, L-arginine and guanidine hydrochloride" 43 : 321-326, 2009

      9 Fu D, "Relationship between thermal inactivation and conformational change of Yarrowia lipolytica lipase and the effect of additives on enzyme stability" 66 : 136-141, 2010

      10 L iu Y, "Purification and characterization of two thermostable glucoamylases produced from Aspergillus niger B -30" 29 : 917-923, 2013

      1 Miller GL, "Use of dinitrosalicylic acid reagent for determination of reducing sugar" 31 : 426-428, 1959

      2 Cui L, "Thermal stability and conformational changes of transglutaminase from a newly isolated Streptomyces hygroscopicus" 99 : 3794-3800, 2008

      3 Ca ves MS, "Thermal inactivation of uricase (urate oxidase): mechanism and effects of additives" 52 : 497-507, 2013

      4 Gangadhara, "The stabilizing effects of polyols and sugars on porcine pancreatic lipase" 86 : 773-781, 2009

      5 Negi S, "Study of conformational changes in glucoamylase of Aspergillus awamori nakazawa in presence of denaturants through CD-spectroscopy" 101 : 7577-7580, 2010

      6 Zaroog MS, "Stabilizing effect of various polyols on the native and the denatured states of glucoamylase" 2013

      7 Chi MC, "Sorbitol counteracts temperature and chemical-induced denaturation of a recombinant α-amylase from alkaliphilic Bacillus sp. TS-23" 39 : 1779-1788, 2012

      8 D ong XY, "Self-interaction of native and denatured lysozyme in the presence of osmolytes, L-arginine and guanidine hydrochloride" 43 : 321-326, 2009

      9 Fu D, "Relationship between thermal inactivation and conformational change of Yarrowia lipolytica lipase and the effect of additives on enzyme stability" 66 : 136-141, 2010

      10 L iu Y, "Purification and characterization of two thermostable glucoamylases produced from Aspergillus niger B -30" 29 : 917-923, 2013

      11 M ichelin M, "Purification and biochemical characterization of a thermostable extracellular glucoamylase produced by the thermotolerant fungus Paecilomyces variotii" 35 : 17-25, 2008

      12 Liu HL, "Protein engineering to improve the thermostability of glucoamylase from Aspergillus awamori based on molecular dynamics simulations" 16 : 19-25, 2003

      13 Rahimzadeh M, "Probing t he role of asparagines mutation in thermostability of Bacillus KR-8104-α-amylase" 50 : 1175-1182, 2012

      14 Devaraneni PK, "Polyolo smolytes stabilize native-like cooperative intermediate state of yeast hexokinase A at low pH" 94 : 947-952, 2012

      15 Sh iba K, "Polydispersity as a parameter for indicating the thermal stability of proteins by dynamic light scattering" 26 : 659-663, 2010

      16 Su RX, "Multilevel structural nature and interactions of bovine serum albumin during heat-induced aggregation process" 22 : 995-1005, 2008

      17 Kumar P, "Microbial glucoamylases:characteristics and applications" 29 : 225-255, 2009

      18 Mertens JA, "Isolation and characterization of a second glucoamylase gene without a starch binding domain from Rhizopus oryzae" 40 : 874-880, 2007

      19 Gangadhara, "Influence of polyols on the stability and kinetic parameters of invertase from Candida utilis: correlation with the conformational stability and activity" 27 : 440-449, 2008

      20 P avezzi FC, "Influence of different substrates on the production of a mutant thermostable glucoamylase in submerged fermentation" 163 : 14-24, 2011

      21 Kell y SM, "How to study proteins by circular dichroism" 1751 : 119-139, 2005

      22 Murayama K, "Heat-induced s econdary structure and conformation change of bovine serum albumin investigated by Fourier transform infrared spectroscopy" 43 : 11526-11532, 2004

      23 Nor ouzian D, "Fungal glucoamylases" 24 : 80-85, 2006

      24 Zaroog MS, "Formation of molten globulelike state during acid denaturation of Aspergillus niger glucoamylase" 47 : 775-784, 2012

      25 Pi ca A, "Exploring the unfolding mechanism of γ-glutamyltranspeptidases: the case of the thermophilic enzyme from Geobacillus thermodenitrificans" 1824 : 571-577, 2012

      26 Az izi A, "Effects of trehalose and sorbitol on the activity and structure of Pseudomonas cepacia lipase: spectroscopic insight" 49 : 652-656, 2011

      27 Ka miyama T, "Effects of modified β-cyclodextrin on thermal stability and conformation of lysozyme" 532 : 10-14, 2012

      28 Sa ntambrogio C, "Effects of methanol on a methanol-tolerant bacterial lipase" 97 : 8609-8618, 2013

      29 Zhang XY, "Effect of microwave irradiation on the structure of glucoamylase" 47 : 2323-2328, 2012

      30 Davis BJ., "Disc electrophoresis II: methods and application to human serum proteins" 121 : 404-427, 1964

      31 Whitmore L, "DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data" 32 : W668-W673, 2004

      32 Au sar SF, "Conformational stability and disassembly of Norwalk virus-like particles: effect of pH and temperature" 281 : 19478-19488, 2006

      33 Zh eng YY, "Cloning, expression, and characterization of a thermostable glucoamylase from Thermoanaerobacter tengcongensis MB4" 87 : 225-233, 2010

      34 Yamamoto T, "Circular dichroism and fluorescence spectroscopy studies of the effect of cyclodextrins on the thermal stability of chicken egg white lysozyme in aqueous solution" 782 : 60-66, 2006

      35 Dock C, "At hermoactive glucoamylase with biotechnological relevance from the thermoacidophilic euryarchaeon Thermoplasma acidophilum" 78 : 105-114, 2008

      36 Johnson WC, "Analyzing protein circular dichroism spectra for accurate secondary structures" 35 : 307-312, 1999

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      기준연도 WOS-KCI 통합IF(2년) KCIF(2년) KCIF(3년)
      2016 1.59 0.33 1.17
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