Bacterial production of poly-γ-glutamic acid and evaluation of its effect on the viability of probiotic microorganisms

2.50
Hdl Handle:
http://hdl.handle.net/2436/241854
Title:
Bacterial production of poly-γ-glutamic acid and evaluation of its effect on the viability of probiotic microorganisms
Authors:
Bhat, Aditya
Abstract:
Poly-γ-glutamic acid (γ-PGA) is a naturally occurring biopolymer made up of repeating units of glutamic acid and can be potentially used for multiple applications. This study compared the production of γ-PGA by eight bacteria (B. subtilis 23856, B. subtilis 23857, B. subtilis 23858 B. subtilis 23859, B. subtilis natto, B. licheniformis 1525, B. licheniformis 6816 and B. licheniformis 9945a) in GS and E media. B. subtilis natto and B. licheniformis 9945a have been investigated extensively for γ-PGA production, however, the remaining six have not previously been used. Using the eight bacteria, yields of up to 22.3 g/l were achieved in shake flasks. On characterization, it was observed that γ-PGA with different properties (crystallinity, acid/salt form and molecular weights ranging from 3,000 Da to 871,000 Da) was produced. Production of γ-PGA by B. subtilis natto in GS medium was scaled up using a fermenter and was tested for novel probiotic applications. The survival of probiotics during freeze drying, storage and ingestion was improved by combining them with a γ-PGA matrix. For L. paracasei, 10% γ-PGA protected the cells significantly better (P < 0.05) than 10% sucrose during freeze drying, whereas for B. longum and B. breve, it showed comparable cryoprotectant activity (P > 0.05) to 10% sucrose. This study also demonstrated the potential use of a non-dairy foodstuff (orange juice) for delivery of probiotics. Two Bifidobacteria strains protected with γ-PGA survived significantly better (P < 0.05) in orange juice for 39 days, with a log reduction in viability of less than 2.99 CFU/ml, when compared to unprotected cells, which showed complete loss in viability by day 20. In addition, γ-PGA protection improved survival of Bifidobacteria in a solution mimicking the environment of the stomach. γ-PGA-protected Bifidobacteria showed little (< 0.47 log CFU/ml) or no loss in viability when stored in simulated gastric juice (pH 2.0) for four hours, whereas unprotected cells died within two hours.
Advisors:
Radecka, Iza Dr
Publisher:
University of Wolverhampton
Issue Date:
Apr-2012
URI:
http://hdl.handle.net/2436/241854
Type:
Thesis or dissertation
Language:
en
Description:
A thesis submitted for the degree of Doctor of Philosophy By Aditya Bhat, MSc
Appears in Collections:
E-Theses

Full metadata record

DC FieldValue Language
dc.contributor.advisorRadecka, Iza Dren_GB
dc.contributor.authorBhat, Adityaen_GB
dc.date.accessioned2012-09-07T13:37:12Z-
dc.date.available2012-09-07T13:37:12Z-
dc.date.issued2012-04-
dc.identifier.urihttp://hdl.handle.net/2436/241854-
dc.descriptionA thesis submitted for the degree of Doctor of Philosophy By Aditya Bhat, MScen_GB
dc.description.abstractPoly-γ-glutamic acid (γ-PGA) is a naturally occurring biopolymer made up of repeating units of glutamic acid and can be potentially used for multiple applications. This study compared the production of γ-PGA by eight bacteria (B. subtilis 23856, B. subtilis 23857, B. subtilis 23858 B. subtilis 23859, B. subtilis natto, B. licheniformis 1525, B. licheniformis 6816 and B. licheniformis 9945a) in GS and E media. B. subtilis natto and B. licheniformis 9945a have been investigated extensively for γ-PGA production, however, the remaining six have not previously been used. Using the eight bacteria, yields of up to 22.3 g/l were achieved in shake flasks. On characterization, it was observed that γ-PGA with different properties (crystallinity, acid/salt form and molecular weights ranging from 3,000 Da to 871,000 Da) was produced. Production of γ-PGA by B. subtilis natto in GS medium was scaled up using a fermenter and was tested for novel probiotic applications. The survival of probiotics during freeze drying, storage and ingestion was improved by combining them with a γ-PGA matrix. For L. paracasei, 10% γ-PGA protected the cells significantly better (P < 0.05) than 10% sucrose during freeze drying, whereas for B. longum and B. breve, it showed comparable cryoprotectant activity (P > 0.05) to 10% sucrose. This study also demonstrated the potential use of a non-dairy foodstuff (orange juice) for delivery of probiotics. Two Bifidobacteria strains protected with γ-PGA survived significantly better (P < 0.05) in orange juice for 39 days, with a log reduction in viability of less than 2.99 CFU/ml, when compared to unprotected cells, which showed complete loss in viability by day 20. In addition, γ-PGA protection improved survival of Bifidobacteria in a solution mimicking the environment of the stomach. γ-PGA-protected Bifidobacteria showed little (< 0.47 log CFU/ml) or no loss in viability when stored in simulated gastric juice (pH 2.0) for four hours, whereas unprotected cells died within two hours.en_GB
dc.language.isoenen
dc.publisherUniversity of Wolverhamptonen
dc.subjectPoly-γ-glutamic aciden_GB
dc.subjectγ-PGAen_GB
dc.subjectbiopolymeren_GB
dc.subjectpolymeren_GB
dc.subjectbiodegradableen_GB
dc.subjectfermentationen_GB
dc.subjectprobioticsen_GB
dc.subjectviabilityen_GB
dc.subjectsurvivalen_GB
dc.titleBacterial production of poly-γ-glutamic acid and evaluation of its effect on the viability of probiotic microorganismsen_GB
dc.typeThesis or dissertationen
dc.rights.embargodate2014-04-
dc.type.qualificationnamePhDen
dc.type.qualificationlevelDoctoralen
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