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Zhang J, Wang L, Luan C, Liu G, Liu J, Zhong Y
Establishment of a rapid and effective plate chromogenic assay for screening of Aspergillus species with high β-fructofuranosidase activity for fructooligosaccharides production
Journal of Microbiological Methods 166 (2019)
ID 105740
Aspergillus niger ATCC 20611
(Ancestor NCBI TaxID 5061,
species name lookup)
Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
NCBI PubMed ID: 31614171Publication DOI: 10.1016/j.mimet.2019.105740Journal NLM ID: 8306883Correspondence: Liu J <liujie

sdu.edu.cn>; Zhong Y <zhongyaohua

sdu.edu.cn>
Institutions: State Key Laboratory of Microbial Technology, Department of Science and Technology Management, Shandong University, Qingdao, China, Zibo Center Hospital, Zi Bo, Shandong Province, China
Fructooligosaccharides (FOS) are commonly regarded as prebiotics and used as components of functional foods. Currently, the industrial sucrose-to-FOS biotransformation is mainly carried out using the microbial-derived β-fructofuranosidases with transglycosylation activity as catalysts. Evaluation of the ability of a microorganism to produce β-fructofuranosidase is commonly conducted by measuring enzyme activity. However, the traditional method requires several steps to identify strains with high β-fructofuranosidase activity, which is not suitable for high-throughput screening. To facilitate screening of a large number of microbial cultures, this study developed a plate chromogenic assay method based on the glucose oxidase (GOD) - peroxidase (POD) bienzymatic system for screening of β-fructofuranosidase-producing fungal strains and predicting their potential to produce FOS. This method used the amount of glucose released from sucrose as indicator to form clear pink halos around the microbial colonies with β-fructofuranosidase activity. Cultivation conditions for the plate assay were optimized as cultivation time 5 h and spore inoculum concentration 100000000 1/ml. Moreover, the method was applied to screening of an Aspergillus niger ATCC 20611 mutant library. The mutant A11 displaying the largest pink halo was screened out and its β-fructofuranosidase activity was determined to be 1.65 fold than that of the parental strain. Thin layer chromatography (TLC) assay further indicated that A11 with the largest halo possessed the highest FOS synthesis ability. These results demonstrated the potential of this plate chromogyenic assay method in the rapid and effective identification of excellent FOS producers from a large number of strain samples.
β-fructofuranosidase, fructooligosaccharides (FOS), plate chromogenic assay, GOD-POD bienzymatic system, Aspergillus niger ATCC 20611
Structure type: oligomer
Location inside paper: Fig. 5, GF2
Trivial name: 1-kestose, inulin
Compound class: fructan, trisaccharide, oligosaccharide, fructo-oligosaccharide
Contained glycoepitopes: IEDB_142488,IEDB_144998,IEDB_146664,IEDB_983931,SB_192
Methods: TLC, cell growth, mutagenesis, enzymatic assay, DNS method, plate chromogenic assay
Enzymes that release or process the structure: β-fructofuranosidase
Synthetic data: enzymatic in vivo
Related record ID(s): 5201, 43612, 44206, 44306, 44492, 45817, 49505, 49521, 49565, 49569, 49572, 49573, 49574, 50321, 50325, 50329, 50332, 50342, 50345, 50348, 50351, 50355, 50358, 50361, 147005, 213017, 219959, 245753, 246867
NCBI Taxonomy refs (TaxIDs): 5061Reference(s) to other database(s): GTC:G11260CN, GlycomeDB:
396, CCSD:
26122, CBank-STR:5624, GenDB:MH445969, US-PT:US2017298332A1
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Zhang J, Wang L, Luan C, Liu G, Liu J, Zhong Y
Establishment of a rapid and effective plate chromogenic assay for screening of Aspergillus species with high β-fructofuranosidase activity for fructooligosaccharides production
Journal of Microbiological Methods 166 (2019)
ID 105740
Aspergillus niger ATCC 20611
(Ancestor NCBI TaxID 5061,
species name lookup)
Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
NCBI PubMed ID: 31614171Publication DOI: 10.1016/j.mimet.2019.105740Journal NLM ID: 8306883Correspondence: Liu J <liujie

sdu.edu.cn>; Zhong Y <zhongyaohua

sdu.edu.cn>
Institutions: State Key Laboratory of Microbial Technology, Department of Science and Technology Management, Shandong University, Qingdao, China, Zibo Center Hospital, Zi Bo, Shandong Province, China
Fructooligosaccharides (FOS) are commonly regarded as prebiotics and used as components of functional foods. Currently, the industrial sucrose-to-FOS biotransformation is mainly carried out using the microbial-derived β-fructofuranosidases with transglycosylation activity as catalysts. Evaluation of the ability of a microorganism to produce β-fructofuranosidase is commonly conducted by measuring enzyme activity. However, the traditional method requires several steps to identify strains with high β-fructofuranosidase activity, which is not suitable for high-throughput screening. To facilitate screening of a large number of microbial cultures, this study developed a plate chromogenic assay method based on the glucose oxidase (GOD) - peroxidase (POD) bienzymatic system for screening of β-fructofuranosidase-producing fungal strains and predicting their potential to produce FOS. This method used the amount of glucose released from sucrose as indicator to form clear pink halos around the microbial colonies with β-fructofuranosidase activity. Cultivation conditions for the plate assay were optimized as cultivation time 5 h and spore inoculum concentration 100000000 1/ml. Moreover, the method was applied to screening of an Aspergillus niger ATCC 20611 mutant library. The mutant A11 displaying the largest pink halo was screened out and its β-fructofuranosidase activity was determined to be 1.65 fold than that of the parental strain. Thin layer chromatography (TLC) assay further indicated that A11 with the largest halo possessed the highest FOS synthesis ability. These results demonstrated the potential of this plate chromogyenic assay method in the rapid and effective identification of excellent FOS producers from a large number of strain samples.
β-fructofuranosidase, fructooligosaccharides (FOS), plate chromogenic assay, GOD-POD bienzymatic system, Aspergillus niger ATCC 20611
Structure type: oligomer
Location inside paper: Fig. 5, GF3
Trivial name: nystose, inulin, 1,1-kestotetraose
Compound class: fructan, tetrasaccharide, oligosaccharide
Contained glycoepitopes: IEDB_142488,IEDB_144998,IEDB_146664,IEDB_923067,IEDB_983931,SB_192
Methods: TLC, cell growth, mutagenesis, enzymatic assay, DNS method, plate chromogenic assay
Enzymes that release or process the structure: β-fructofuranosidase
Synthetic data: enzymatic in vivo
Related record ID(s): 5421, 43613, 44207, 44307, 44493, 49506, 49522, 49571, 49573, 49575, 50322, 50326, 50330, 50333, 50343, 50346, 50349, 50352, 50356, 50359, 50362, 100071, 147006, 244675, 269959
NCBI Taxonomy refs (TaxIDs): 5061Reference(s) to other database(s): GTC:G60726JL, GlycomeDB:
63, CCSD:
23519, CBank-STR:9595, GenDB:MH445969, US-PT:US2017298332A1
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Zhang J, Wang L, Luan C, Liu G, Liu J, Zhong Y
Establishment of a rapid and effective plate chromogenic assay for screening of Aspergillus species with high β-fructofuranosidase activity for fructooligosaccharides production
Journal of Microbiological Methods 166 (2019)
ID 105740
Aspergillus niger ATCC 20611
(Ancestor NCBI TaxID 5061,
species name lookup)
Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
NCBI PubMed ID: 31614171Publication DOI: 10.1016/j.mimet.2019.105740Journal NLM ID: 8306883Correspondence: Liu J <liujie

sdu.edu.cn>; Zhong Y <zhongyaohua

sdu.edu.cn>
Institutions: State Key Laboratory of Microbial Technology, Department of Science and Technology Management, Shandong University, Qingdao, China, Zibo Center Hospital, Zi Bo, Shandong Province, China
Fructooligosaccharides (FOS) are commonly regarded as prebiotics and used as components of functional foods. Currently, the industrial sucrose-to-FOS biotransformation is mainly carried out using the microbial-derived β-fructofuranosidases with transglycosylation activity as catalysts. Evaluation of the ability of a microorganism to produce β-fructofuranosidase is commonly conducted by measuring enzyme activity. However, the traditional method requires several steps to identify strains with high β-fructofuranosidase activity, which is not suitable for high-throughput screening. To facilitate screening of a large number of microbial cultures, this study developed a plate chromogenic assay method based on the glucose oxidase (GOD) - peroxidase (POD) bienzymatic system for screening of β-fructofuranosidase-producing fungal strains and predicting their potential to produce FOS. This method used the amount of glucose released from sucrose as indicator to form clear pink halos around the microbial colonies with β-fructofuranosidase activity. Cultivation conditions for the plate assay were optimized as cultivation time 5 h and spore inoculum concentration 100000000 1/ml. Moreover, the method was applied to screening of an Aspergillus niger ATCC 20611 mutant library. The mutant A11 displaying the largest pink halo was screened out and its β-fructofuranosidase activity was determined to be 1.65 fold than that of the parental strain. Thin layer chromatography (TLC) assay further indicated that A11 with the largest halo possessed the highest FOS synthesis ability. These results demonstrated the potential of this plate chromogyenic assay method in the rapid and effective identification of excellent FOS producers from a large number of strain samples.
β-fructofuranosidase, fructooligosaccharides (FOS), plate chromogenic assay, GOD-POD bienzymatic system, Aspergillus niger ATCC 20611
Structure type: oligomer
Location inside paper: Fig. 5, GF4
Trivial name: 1-fructofuranosylsylnystose, 1-fructofuranosylnystose
Compound class: pentasaccharide, oligosaccharide
Contained glycoepitopes: IEDB_142488,IEDB_144998,IEDB_146664,IEDB_923067,IEDB_983931,SB_192
Methods: TLC, cell growth, mutagenesis, enzymatic assay, DNS method, plate chromogenic assay
Enzymes that release or process the structure: β-fructofuranosidase
Synthetic data: enzymatic in vivo
Related record ID(s): 44208, 44308, 49507, 49571, 49572, 49576, 50323, 50331, 50344, 50347, 50350, 50353, 50357, 50360, 50363, 269960
NCBI Taxonomy refs (TaxIDs): 5061Reference(s) to other database(s): GenDB:MH445969, US-PT:US2017298332A1
Show glycosyltransferases
There is only one chemically distinct structure:
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