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Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis

Medium chain length poly-3-hydroxyalkanoates (mcl-PHA) is a biodegradable polymer made of different hydroxyalkanoates with (R)-configuration as monomeric units and a source of valuable biomaterials. Biosynthesis, in vivo and in vitro depolymerization of intracellular mcl-PHA in Pseudomonas putida...

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Main Author:	Siti Nor Syairah , Anis
Format:	Thesis
Published:	2017
Subjects:	Q Science (General) QD Chemistry
Online Access:	http://studentsrepo.um.edu.my/8132/1/All.pdf http://studentsrepo.um.edu.my/8132/6/syairah.pdf http://studentsrepo.um.edu.my/8132/
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spelling	my.um.stud.81322020-09-07T20:35:35Z Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis Siti Nor Syairah , Anis Q Science (General) QD Chemistry Medium chain length poly-3-hydroxyalkanoates (mcl-PHA) is a biodegradable polymer made of different hydroxyalkanoates with (R)-configuration as monomeric units and a source of valuable biomaterials. Biosynthesis, in vivo and in vitro depolymerization of intracellular mcl-PHA in Pseudomonas putida Bet001 grown on lauric acid was studied. Highest mcl-PHA fraction (> 50 % of total biomass) and cell concentration (8 g L-1) were obtained at carbon-to-nitrogen (C/N) ratio 20, initial cell concentration 1 g L-1, and 48 hours fermentation. The mcl-PHA comprised of 3-hydroxyhexanoate (C6) (6.1 ± 2.0 mol%), 3-hydroxyoctanote (C8) (40.1 ± 2.7 mol%), 3-hydroxydecanoate (C10) (32.8 ± 2.8 mol%) and 3-hydroxydodecanoate (C12) (21.0 ± 3.2 mol%). In vivo action was studied in a mineral liquid medium without carbon source, and in different buffer solutions with varied pH, molarity, ionic strength (I) and temperature. Rate and percentage of in vivo depolymerization were highest in 0.2 M Tris-HCl buffer (pH 9, I = 0.2 M, 30 ºC) at 0.21 g L-1 h-1 and 98.6 ± 1.3 wt%, respectively. There is a congruity visà- vis to specific buffer type, molarity, pH, I and temperature values for superior in vivo depolymerization activities. For in vitro depolymerization studies, cell-free system was obtained from lysing bacterial cells suspension by ultrasonication at optimum conditions (frequency 37 kHz, 30 % of power output, < 25 ºC for 120 minutes). The comparison between in vivo and in vitro depolymerizations of intracellular mcl-PHA was made. In vitro depolymerization showed lower depolymerization rates and activities compared to in vivo depolymerization. However, final product yield of in vitro depolymerization showed higher yield compared to in vivo depolymerization. The monomer liberation rate reflected the mol% distribution of the initial polymer subunit composition, and the resulting direct individual products of depolymerization were identical for both in vivo and in vitro processes. It points to exo-type reaction for both processes, and potential biological route to chiral molecules. 2017-09 Thesis NonPeerReviewed application/pdf http://studentsrepo.um.edu.my/8132/1/All.pdf application/pdf http://studentsrepo.um.edu.my/8132/6/syairah.pdf Siti Nor Syairah , Anis (2017) Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis. PhD thesis, University of Malaya. http://studentsrepo.um.edu.my/8132/
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Student Repository
url_provider	http://studentsrepo.um.edu.my/
topic	Q Science (General) QD Chemistry
spellingShingle	Q Science (General) QD Chemistry Siti Nor Syairah , Anis Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
description	Medium chain length poly-3-hydroxyalkanoates (mcl-PHA) is a biodegradable polymer made of different hydroxyalkanoates with (R)-configuration as monomeric units and a source of valuable biomaterials. Biosynthesis, in vivo and in vitro depolymerization of intracellular mcl-PHA in Pseudomonas putida Bet001 grown on lauric acid was studied. Highest mcl-PHA fraction (> 50 % of total biomass) and cell concentration (8 g L-1) were obtained at carbon-to-nitrogen (C/N) ratio 20, initial cell concentration 1 g L-1, and 48 hours fermentation. The mcl-PHA comprised of 3-hydroxyhexanoate (C6) (6.1 ± 2.0 mol%), 3-hydroxyoctanote (C8) (40.1 ± 2.7 mol%), 3-hydroxydecanoate (C10) (32.8 ± 2.8 mol%) and 3-hydroxydodecanoate (C12) (21.0 ± 3.2 mol%). In vivo action was studied in a mineral liquid medium without carbon source, and in different buffer solutions with varied pH, molarity, ionic strength (I) and temperature. Rate and percentage of in vivo depolymerization were highest in 0.2 M Tris-HCl buffer (pH 9, I = 0.2 M, 30 ºC) at 0.21 g L-1 h-1 and 98.6 ± 1.3 wt%, respectively. There is a congruity visà- vis to specific buffer type, molarity, pH, I and temperature values for superior in vivo depolymerization activities. For in vitro depolymerization studies, cell-free system was obtained from lysing bacterial cells suspension by ultrasonication at optimum conditions (frequency 37 kHz, 30 % of power output, < 25 ºC for 120 minutes). The comparison between in vivo and in vitro depolymerizations of intracellular mcl-PHA was made. In vitro depolymerization showed lower depolymerization rates and activities compared to in vivo depolymerization. However, final product yield of in vitro depolymerization showed higher yield compared to in vivo depolymerization. The monomer liberation rate reflected the mol% distribution of the initial polymer subunit composition, and the resulting direct individual products of depolymerization were identical for both in vivo and in vitro processes. It points to exo-type reaction for both processes, and potential biological route to chiral molecules.
format	Thesis
author	Siti Nor Syairah , Anis
author_facet	Siti Nor Syairah , Anis
author_sort	Siti Nor Syairah , Anis
title	Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
title_short	Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
title_full	Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
title_fullStr	Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
title_full_unstemmed	Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis
title_sort	biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / siti nor syairah anis
publishDate	2017
url	http://studentsrepo.um.edu.my/8132/1/All.pdf http://studentsrepo.um.edu.my/8132/6/syairah.pdf http://studentsrepo.um.edu.my/8132/
_version_	1738506105940606976
score	13.211869

Biosynthesis, in vivo and in vitro depolymerizations of intracellular medium chain length poly-3 hydroxyalkanoates / Siti Nor Syairah Anis

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