79c214001e08a1e5f899111b175760b2  metabolites_2014v4p281.pdf
9316c2c2886dfd8bca6c6953d1cddb10f532ad30  metabolites_2014v4p281.pdf
29df5433388e43bf4eb4e108fac12add0465cab728a6f539220d894a1dcdee32  metabolites_2014v4p281.pdf

Title:          Quantitative Metabolomics and Instationary 13C-Metabolic Flux Analysis Reveals Impact of Recombinant Protein Production on Trehalose and Energy Metabolism in Pichia pastoris
Subject:        Pichia pastoris has been recognized as an effective host for recombinant protein production. In this work, we combine metabolomics and instationary 13C metabolic flux analysis (INST 13C-MFA) using GC-MS and LC-MS/MS to evaluate the potential impact of the production of a Rhizopus oryzae lipase (Rol) on P. pastoris central carbon metabolism. Higher oxygen uptake and CO2 production rates and slightly reduced biomass yield suggest an increased energy demand for the producing strain. This observation is further confirmed by 13C-based metabolic flux analysis. In particular, the flux through the methanol oxidation pathway and the TCA cycle was increased in the Rol-producing strain compared to the reference strain. Next to changes in the flux distribution, significant variations in intracellular metabolite concentrations were observed. Most notably, the pools of trehalose, which is related to cellular stress response, and xylose, which is linked to methanol assimilation, were significantly increased in the recombinant strain.
Keywords:       Pichia pastoris; instationary 13C-metabolic flux analysis; recombinant protein production; trehalose; metabolome; GC-MS; LC-MS
Author:         Joel Jordà, Hugo Cueto Rojas, Marc Carnicer, Aljoscha Wahl, Pau Ferrer, Joan Albiol
Creator:        PScript5.dll Version 5.2.2
Producer:       Acrobat Distiller 9.0.0 (Windows)
CreationDate:   Mon May  5 17:31:15 2014
ModDate:        Fri Feb 19 12:17:18 2016
Tagged:         no
UserProperties: no
Suspects:       no
Form:           none
JavaScript:     no
Pages:          19
Encrypted:      no
Page size:      595.22 x 842 pts (A4)
Page rot:       0
File size:      504609 bytes
Optimized:      yes
PDF version:    1.5

name                                 type              encoding         emb sub uni object ID
------------------------------------ ----------------- ---------------- --- --- --- ---------
PFJGBD+TimesNewRoman,Italic          TrueType          WinAnsi          yes yes no     171  0
PFJGDE+TimesNewRoman,Bold            TrueType          WinAnsi          yes yes no     173  0
PFJGFF+TimesNewRoman                 TrueType          WinAnsi          yes yes no     175  0
PFJGHH+TimesNewRoman,BoldItalic      TrueType          WinAnsi          yes yes no     177  0
PFJGFE+TimesNewRoman                 CID TrueType      Identity-H       yes yes yes    139  0
PFJJCG+Symbol                        CID TrueType      Identity-H       yes yes yes    138  0
PFJLOG+Arial                         TrueType          WinAnsi          yes yes no     145  0
PFJLPI+Arial,Bold                    TrueType          WinAnsi          yes yes no     143  0
PFJMAI+Wingdings2                    CID TrueType      Identity-H       yes yes yes    142  0
PFJODM+Calibri,Bold                  TrueType          WinAnsi          yes yes no     148  0

Jhove (Rel. 1.6, 2011-01-04)
 Date: 2016-02-24 08:03:50 CET
 RepresentationInformation: metabolites_2014v4p281.pdf
  ReportingModule: BYTESTREAM, Rel. 1.3 (2007-04-10)
  LastModified: 2016-02-19 12:17:38 CET
  Size: 504609
  Format: bytestream
  Status: Well-Formed and valid
  SignatureMatches:
   PDF-hul
  MIMEtype: application/octet-stream
  Checksum: 6f8d167a
   Type: CRC32
  Checksum: 79c214001e08a1e5f899111b175760b2
   Type: MD5
  Checksum: 9316c2c2886dfd8bca6c6953d1cddb10f532ad30
   Type: SHA-1