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Coexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri

A multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade beta -glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001... Full description

Journal Title: International Journal of Molecular Sciences 0, 2014, Vol.15(7), pp.12842-12860
Main Author: Wang, Lei
Other Authors: Yang, Yuxin , Cai, Bei , Cao, Pinghua , Yang, Mingming , Chen, Yulin
Format: Electronic Article Electronic Article
Language: English
Subjects:
Age
ID: ISSN: 1422-0067 ; DOI: 10.3390/ijms150712842
Link: http://search.proquest.com/docview/1727672185/?pq-origsite=primo
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title: Coexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri
format: Article
creator:
  • Wang, Lei
  • Yang, Yuxin
  • Cai, Bei
  • Cao, Pinghua
  • Yang, Mingming
  • Chen, Yulin
subjects:
  • Endoglucanase
  • Feeding
  • Age
  • Poultry
  • Calcium
  • Secretion
  • Phosphorus
  • Probiotics
  • Enzymes
  • Phytic Acid
  • Nutrient Utilization
  • Gel Electrophoresis
  • Supplementation
  • L-Lactate Dehydrogenase
  • Expression Vectors
  • Beta -Glucan
  • Promoters
  • 6-Phytase
  • Secretion Signals
  • Molecular Weight
  • Cecum
  • Gastrointestinal Tract
  • Body Weight Gain
  • Lactobacillus Casei
  • Lactococcus Lactis
  • Lactobacillus Reuteri
  • Bacillus Subtilis
  • Bacteroides Vulgatus
  • Aspergillus Fumigatus
  • Veillonella
  • Escherichia Coli
  • Bifidobacterium
  • Human Diseases
  • Plants and Algae
  • Coexpression
  • Secretion
  • Endoglucanase
  • Phytase
  • Lactobacillus Reuteri
  • Broiler
ispartof: International Journal of Molecular Sciences, 0, 2014, Vol.15(7), pp.12842-12860
description: A multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade beta -glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001 endoglucanase gene (celW) and Aspergillus fumigatus WL002 phytase gene (phyW) mature peptide (phyWM) were cloned into an expression vector with the lactate dehydrogenase promoter of Lactobacillus casei and the secretion signal peptide of the Lactococcus lactis usp45 gene. This construct was then transformed into Lactobacillus reuteri XC1 that had been isolated from the gastrointestinal tract of broilers. Heterologous enzyme production and feed effectiveness of this genetically modified L. reuteri strain were investigated and evaluated. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the molecular mass of phyWM and celW was approximately 48.2 and 55 kDa, respectively, consistent with their predicted molecular weights. Endoglucanase and phytase activities in the extracellular fraction of the transformed L. reuteri culture were 0.68 and 0.42 U/mL, respectively. Transformed L. reuteri improved the feed conversion ratio of broilers from 21 to 42 days of age and over the whole feeding period. However, there was no effect on body weight gain and feed intake of chicks. Transformed L. reuteri supplementation improved levels of ash, calcium and phosphorus in tibiae at day 21 and of phosphorus at day 42. In addition, populations of Escherichia coli, Veillonella spp. and Bacteroides vulgatus were decreased, while populations of Bifidobacterium genus and Lactobacillus spp. were increased in the cecum at day 21.
language: eng
source:
identifier: ISSN: 1422-0067 ; DOI: 10.3390/ijms150712842
fulltext: fulltext
issn:
  • 14220067
  • 1422-0067
url: Link


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titleCoexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri
creatorWang, Lei ; Yang, Yuxin ; Cai, Bei ; Cao, Pinghua ; Yang, Mingming ; Chen, Yulin
contributorWang, Lei (correspondence author)
ispartofInternational Journal of Molecular Sciences, 0, 2014, Vol.15(7), pp.12842-12860
identifierISSN: 1422-0067 ; DOI: 10.3390/ijms150712842
subjectEndoglucanase ; Feeding ; Age ; Poultry ; Calcium ; Secretion ; Phosphorus ; Probiotics ; Enzymes ; Phytic Acid ; Nutrient Utilization ; Gel Electrophoresis ; Supplementation ; L-Lactate Dehydrogenase ; Expression Vectors ; Beta -Glucan ; Promoters ; 6-Phytase ; Secretion Signals ; Molecular Weight ; Cecum ; Gastrointestinal Tract ; Body Weight Gain ; Lactobacillus Casei ; Lactococcus Lactis ; Lactobacillus Reuteri ; Bacillus Subtilis ; Bacteroides Vulgatus ; Aspergillus Fumigatus ; Veillonella ; Escherichia Coli ; Bifidobacterium ; Human Diseases ; Plants and Algae ; Coexpression ; Secretion ; Endoglucanase ; Phytase ; Lactobacillus Reuteri ; Broiler
descriptionA multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade beta -glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001 endoglucanase gene (celW) and Aspergillus fumigatus WL002 phytase gene (phyW) mature peptide (phyWM) were cloned into an expression vector with the lactate dehydrogenase promoter of Lactobacillus casei and the secretion signal peptide of the Lactococcus lactis usp45 gene. This construct was then transformed into Lactobacillus reuteri XC1 that had been isolated from the gastrointestinal tract of broilers. Heterologous enzyme production and feed effectiveness of this genetically modified L. reuteri strain were investigated and evaluated. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the molecular mass of phyWM and celW was approximately 48.2 and 55 kDa, respectively, consistent with their predicted molecular weights. Endoglucanase and phytase activities in the extracellular fraction of the transformed L. reuteri culture were 0.68 and 0.42 U/mL, respectively. Transformed L. reuteri improved the feed conversion ratio of broilers from 21 to 42 days of age and over the whole feeding period. However, there was no effect on body weight gain and feed intake of chicks. Transformed L. reuteri supplementation improved levels of ash, calcium and phosphorus in tibiae at day 21 and of phosphorus at day 42. In addition, populations of Escherichia coli, Veillonella spp. and Bacteroides vulgatus were decreased, while populations of Bifidobacterium genus and Lactobacillus spp. were increased in the cecum at day 21.
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titleCoexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri
descriptionA multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade beta -glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001 endoglucanase gene (celW) and Aspergillus fumigatus WL002 phytase gene (phyW) mature peptide (phyWM) were cloned into an expression vector with the lactate dehydrogenase promoter of Lactobacillus casei and the secretion signal peptide of the Lactococcus lactis usp45 gene. This construct was then transformed into Lactobacillus reuteri XC1 that had been isolated from the gastrointestinal tract of broilers. Heterologous enzyme production and feed effectiveness of this genetically modified L. reuteri strain were investigated and evaluated. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the molecular mass of phyWM and celW was approximately 48.2 and 55 kDa, respectively, consistent with their predicted molecular weights. Endoglucanase and phytase activities in the extracellular fraction of the transformed L. reuteri culture were 0.68 and 0.42 U/mL, respectively. Transformed L. reuteri improved the feed conversion ratio of broilers from 21 to 42 days of age and over the whole feeding period. However, there was no effect on body weight gain and feed intake of chicks. Transformed L. reuteri supplementation improved levels of ash, calcium and phosphorus in tibiae at day 21 and of phosphorus at day 42. In addition, populations of Escherichia coli, Veillonella spp. and Bacteroides vulgatus were decreased, while populations of Bifidobacterium genus and Lactobacillus spp. were increased in the cecum at day 21.
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8Enzymes
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10Nutrient Utilization
11Gel Electrophoresis
12Supplementation
13L-Lactate Dehydrogenase
14Expression Vectors
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16Promoters
176-Phytase
18Secretion Signals
19Molecular Weight
20Cecum
21Gastrointestinal Tract
22Body Weight Gain
23Lactobacillus Casei
24Lactococcus Lactis
25Lactobacillus Reuteri
26Bacillus Subtilis
27Bacteroides Vulgatus
28Aspergillus Fumigatus
29Veillonella
30Escherichia Coli
31Bifidobacterium
32Human Diseases
33Plants and Algae
34Coexpression
35Phytase
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titleCoexpression and Secretion of Endoglucanase and Phytase Genes in Lactobacillus reuteri
authorWang, Lei ; Yang, Yuxin ; Cai, Bei ; Cao, Pinghua ; Yang, Mingming ; Chen, Yulin
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abstractA multifunctional transgenic Lactobacillus with probiotic characteristics and an ability to degrade beta -glucan and phytic acid (phytate) was engineered to improve nutrient utilization, increase production performance and decrease digestive diseases in broiler chickens. The Bacillus subtilis WL001 endoglucanase gene (celW) and Aspergillus fumigatus WL002 phytase gene (phyW) mature peptide (phyWM) were cloned into an expression vector with the lactate dehydrogenase promoter of Lactobacillus casei and the secretion signal peptide of the Lactococcus lactis usp45 gene. This construct was then transformed into Lactobacillus reuteri XC1 that had been isolated from the gastrointestinal tract of broilers. Heterologous enzyme production and feed effectiveness of this genetically modified L. reuteri strain were investigated and evaluated. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed that the molecular mass of phyWM and celW was approximately 48.2 and 55 kDa, respectively, consistent with their predicted molecular weights. Endoglucanase and phytase activities in the extracellular fraction of the transformed L. reuteri culture were 0.68 and 0.42 U/mL, respectively. Transformed L. reuteri improved the feed conversion ratio of broilers from 21 to 42 days of age and over the whole feeding period. However, there was no effect on body weight gain and feed intake of chicks. Transformed L. reuteri supplementation improved levels of ash, calcium and phosphorus in tibiae at day 21 and of phosphorus at day 42. In addition, populations of Escherichia coli, Veillonella spp. and Bacteroides vulgatus were decreased, while populations of Bifidobacterium genus and Lactobacillus spp. were increased in the cecum at day 21.
doi10.3390/ijms150712842
urlhttp://search.proquest.com/docview/1727672185/
date2014-01-01