Metabolic Flux Analysis for Lipid Yield Improvement in Mucor rouxii ATCC24905

การวิเคราะห์การกระจายตัวของเมตาโบลิคฟลักซ์ในรา Mucor rouxii ATCC24905 เพื่อหาชุมทางหลักในการผลิตไขมัน

Name: Phichit Assawachatch

Organization : มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Name: พิชิต อัศวชาติชาญชัย

keyword: Lipid Yield

; Mucor rouxii

; Metabolic Flux Analysis

; Principal Nodes

; Sensitivity Analysis

Abstract: Mucor rouxii ATCC24905, a filamentous fungus, is a rich source of gamma linolenic acid (GLA) which is of great importance because of its pharmaceutical properties such as preventing heart diseases. The GLA proportion in the lipid produced by filamentous Mucor species is very high. The higher GLA proportion is also found in the culture grown at lower temperature. In this study, metabolic flux analysis was applied to determine the cellular carbon flux distributions in a simplified metabolic network of Mucor rouxii ATCC24905 during aerobic growth at 15 ?C and 35 ?C on glucose–limited chemostat culture . A simplified metabolic network of M. rouxii was first constructed primarily from the existing biochemical reactions of Aspergillus oryzae and Saccharomyces cerevisiae. The metabolic pathways include transport reactions, catabolism: EMP, pentose phosphate pathways, TCA cycle, anaplerotic reaction and biosynthesis reactions. Then, a stoichiometric model reflecting the pathways was setup. The stoichiometric network consists of 70 pathway reactions and 63 intracellular metabolites, resulting in a system having a degree of freedom of 7. To successfully quantify the internal metabolic fluxes by applying metabolite balances around the pseudo- steady state assumed intracellular metabolites, we experimentally measured the specific consumption rates of glucose and ammonia, the specific secretion rate of ethanol, and the specific formation rates of lipid and protein. Besides, we adopted the specific formation rates of DNA and RNA from A. oryzae to fulfill of the freedom. The importance of basing the flux analysis on accurate measurements was demonstrated through a sensitivity analysis. According to sensitivity analysis, the condition number of this model was 1142. Hence, the calculation of intracellular fluxes was fairly accurate. It was also found that the accuracy of the measurements of specific lipid production rate was critical for the correct calculation of the flux distribution. The flux split ratio analysis indicated that there are three principal nodes affecting lipid yield of M. rouxii ATCC24905: glucose 6-phosphate node, pyruvate node and acetaldehyde node

มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: Lect.Asawin Meechai

Role: Supervisor

Created: 2002

Issued: 2005-10-03

Modified: 2006-05-10

วิทยานิพนธ์/Thesis

ISBN: 9749551184

CallNumber: BIT932

tha

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biotechnology

Grantor: King Mongkut's University of Technology Thonburi

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