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Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase

Fructooligosaccharides (FOSs)—fructose-based oligosaccharides—are typical prebiotics with health-promoting effects in humans and animals. The trisaccharide 1-kestotriose is the most attractive inulin-type FOS. We previously reported a recombinant sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4...

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Autores principales: Pérez, Enrique R, Martínez, Duniesky, Menéndez, Carmen, Alfonso, Dubiel, Rodríguez, Iván, Trujillo, Luis E, Sobrino, Alina, Ramírez, Ricardo, Pimentel, Eulogio, Hernández, Lázaro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9113426/
https://www.ncbi.nlm.nih.gov/pubmed/34137896
http://dx.doi.org/10.1093/jimb/kuab036
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author Pérez, Enrique R
Martínez, Duniesky
Menéndez, Carmen
Alfonso, Dubiel
Rodríguez, Iván
Trujillo, Luis E
Sobrino, Alina
Ramírez, Ricardo
Pimentel, Eulogio
Hernández, Lázaro
author_facet Pérez, Enrique R
Martínez, Duniesky
Menéndez, Carmen
Alfonso, Dubiel
Rodríguez, Iván
Trujillo, Luis E
Sobrino, Alina
Ramírez, Ricardo
Pimentel, Eulogio
Hernández, Lázaro
author_sort Pérez, Enrique R
collection PubMed
description Fructooligosaccharides (FOSs)—fructose-based oligosaccharides—are typical prebiotics with health-promoting effects in humans and animals. The trisaccharide 1-kestotriose is the most attractive inulin-type FOS. We previously reported a recombinant sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99) from Schedonorus arundinaceus (Sa) that efficiently converts sucrose into 1-kestotriose. In this study, Pichia pastoris PGFT6x-308 constitutively expressing nine copies of the Sa1-SST gene displayed fructosyltransferase activity in undisrupted biomass (49.8 U/ml) and culture supernatant (120.7 U/ml) in fed-batch fermentation (72 hr) with sugarcane molasses. Toluene permeabilization increased 2.3-fold the Sa1-SSTrec activity of whole cells entrapped in calcium-alginate beads. The reaction with refined or raw sugar (600 g/l) yielded 1-kestotriose and 1,1-kestotetraose in a ratio of 8:2 with their sum representing above 55% (wt/wt) of total carbohydrates. The FOSs yield decreased to 45% (wt/wt) when sugarcane syrup and molasses were used as cheaper sucrose sources. The beads retained 80% residual Sa1-SSTrec activity after a 30-day batchwise operation with refined cane sugar at 30°C and pH 5.5. The immobilized biocatalyst is attractive for the continuous production of short-chain FOSs, most particularly 1-kestotriose.
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spelling pubmed-91134262022-06-08 Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase Pérez, Enrique R Martínez, Duniesky Menéndez, Carmen Alfonso, Dubiel Rodríguez, Iván Trujillo, Luis E Sobrino, Alina Ramírez, Ricardo Pimentel, Eulogio Hernández, Lázaro J Ind Microbiol Biotechnol Biocatalysis Fructooligosaccharides (FOSs)—fructose-based oligosaccharides—are typical prebiotics with health-promoting effects in humans and animals. The trisaccharide 1-kestotriose is the most attractive inulin-type FOS. We previously reported a recombinant sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99) from Schedonorus arundinaceus (Sa) that efficiently converts sucrose into 1-kestotriose. In this study, Pichia pastoris PGFT6x-308 constitutively expressing nine copies of the Sa1-SST gene displayed fructosyltransferase activity in undisrupted biomass (49.8 U/ml) and culture supernatant (120.7 U/ml) in fed-batch fermentation (72 hr) with sugarcane molasses. Toluene permeabilization increased 2.3-fold the Sa1-SSTrec activity of whole cells entrapped in calcium-alginate beads. The reaction with refined or raw sugar (600 g/l) yielded 1-kestotriose and 1,1-kestotetraose in a ratio of 8:2 with their sum representing above 55% (wt/wt) of total carbohydrates. The FOSs yield decreased to 45% (wt/wt) when sugarcane syrup and molasses were used as cheaper sucrose sources. The beads retained 80% residual Sa1-SSTrec activity after a 30-day batchwise operation with refined cane sugar at 30°C and pH 5.5. The immobilized biocatalyst is attractive for the continuous production of short-chain FOSs, most particularly 1-kestotriose. Oxford University Press 2021-06-17 /pmc/articles/PMC9113426/ /pubmed/34137896 http://dx.doi.org/10.1093/jimb/kuab036 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Society of Industrial Microbiology and Biotechnology. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Biocatalysis
Pérez, Enrique R
Martínez, Duniesky
Menéndez, Carmen
Alfonso, Dubiel
Rodríguez, Iván
Trujillo, Luis E
Sobrino, Alina
Ramírez, Ricardo
Pimentel, Eulogio
Hernández, Lázaro
Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title_full Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title_fullStr Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title_full_unstemmed Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title_short Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
title_sort fructooligosaccharides production by immobilized pichia pastoris cells expressing schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase
topic Biocatalysis
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9113426/
https://www.ncbi.nlm.nih.gov/pubmed/34137896
http://dx.doi.org/10.1093/jimb/kuab036
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