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Introduction and characterization of...

Hahn, J. J.

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Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures./
Author:	Hahn, J. J.
Description:	248 p.
Notes:	Adviser: Friedrich Srienc.
Contained By:	Dissertation Abstracts International59-03B.
Subject:	Agriculture, Plant Culture. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9826830
ISBN:	0591792664

Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures.
Hahn, J. J.

Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures. - 248 p.

Adviser: Friedrich Srienc.

Thesis (Ph.D.)--University of Minnesota, 1998.

The expression in plants of biosynthetic pathways leading to the production of the commercially-valuable biodegradable polymer poly(3-hydroxybutyrate) (PHB) was investigated. For this purpose, the growth kinetics and nutrient uptake of Black Mexican Sweet (BMS) maize cell cultures (Zea mays L.) were thoroughly characterized both in a 2 L modified mammalian cell bioreactor and in shake flasks. Expression of the first two genes of the Ralstonia eutropha PHB biosynthetic pathway, encoding $\beta$-ketothiolase and acetoacetyl-CoA reductase, led to a slight increase in PHB precursor production and a significant decrease in growth rate. Each of the three genes of the Ralstonia eutropha PHB biosynthetic pathway was modified by the addition of a six amino acid transit peptide to the carboxy terminus in order to target expression to the peroxisome. This resulted in up to 0.4 mg/g fresh weight PHB when only the final gene of the pathway, encoding the PHA synthase enzyme, was expressed and up to 2 mg/g fresh weight PHB when all three genes of the pathway were expressed. In both cases, a slight decrease in culture growth rates was observed. The peroxisomal localization of PHB accumulation was verified by means of sucrose density gradient fractionation and freeze-fracture electron microscopy. Additional work involved the expression of the Pseudomonas oleovorans PHA Polymerase 2 gene in maize peroxisomes and the expression of the R. eutropha PHA synthase gene in the peroxisomes of intact tobacco and Arabidopsis thaliana plants. In tobacco (Nicotiana tabacum) plants, preliminary gas chromatography measurements indicate accumulation of PHB in leaves and roots of up to 3% of dry weight and in senescent leaves of up to 5% of dry weight. Metabolic modeling was used to predict PHBV copolymer composition, and a preliminary cost analysis was developed for a plant to co-produce corn starch and PHB from transgenic maize.

ISBN: 0591792664Subjects--Topical Terms:

1018669
Agriculture, Plant Culture.

Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures.
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005 20110426
008 110426s1998 eng d
020 $a 0591792664
035 $a (UnM)AAI9826830
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100 1 $a Hahn, J. J. $3 1251798
245 1 0 $a Introduction and characterization of the poly(3-hydroxybutyrate) biosynthetic pathway in plant cell cultures.
300 $a 248 p.
500 $a Adviser: Friedrich Srienc.
500 $a Source: Dissertation Abstracts International, Volume: 59-03, Section: B, page: 1231.
502 $a Thesis (Ph.D.)--University of Minnesota, 1998.
520 $a The expression in plants of biosynthetic pathways leading to the production of the commercially-valuable biodegradable polymer poly(3-hydroxybutyrate) (PHB) was investigated. For this purpose, the growth kinetics and nutrient uptake of Black Mexican Sweet (BMS) maize cell cultures (Zea mays L.) were thoroughly characterized both in a 2 L modified mammalian cell bioreactor and in shake flasks. Expression of the first two genes of the Ralstonia eutropha PHB biosynthetic pathway, encoding $\beta$-ketothiolase and acetoacetyl-CoA reductase, led to a slight increase in PHB precursor production and a significant decrease in growth rate. Each of the three genes of the Ralstonia eutropha PHB biosynthetic pathway was modified by the addition of a six amino acid transit peptide to the carboxy terminus in order to target expression to the peroxisome. This resulted in up to 0.4 mg/g fresh weight PHB when only the final gene of the pathway, encoding the PHA synthase enzyme, was expressed and up to 2 mg/g fresh weight PHB when all three genes of the pathway were expressed. In both cases, a slight decrease in culture growth rates was observed. The peroxisomal localization of PHB accumulation was verified by means of sucrose density gradient fractionation and freeze-fracture electron microscopy. Additional work involved the expression of the Pseudomonas oleovorans PHA Polymerase 2 gene in maize peroxisomes and the expression of the R. eutropha PHA synthase gene in the peroxisomes of intact tobacco and Arabidopsis thaliana plants. In tobacco (Nicotiana tabacum) plants, preliminary gas chromatography measurements indicate accumulation of PHB in leaves and roots of up to 3% of dry weight and in senescent leaves of up to 5% of dry weight. Metabolic modeling was used to predict PHBV copolymer composition, and a preliminary cost analysis was developed for a plant to co-produce corn starch and PHB from transgenic maize.
590 $a School code: 0130.
650 4 $a Agriculture, Plant Culture. $3 1018669
650 4 $a Biology, Molecular. $3 1017719
650 4 $a Engineering, Chemical. $3 1018531
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710 2 0 $a University of Minnesota. $3 676231
773 0 $t Dissertation Abstracts International $g 59-03B.
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792 $a 1998
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9826830