High-level expression of Rhodotorula gracilis d-amino acid oxidase in Pichia pastoris

被引：12
作者：
Abad, Sandra [1 ]
Nahalka, Jozef [1 ,2 ]
Winkler, Margit [1 ]
Bergler, Gabriele [1 ]
Speight, Robert [4 ]
Glieder, Anton [1 ,3 ]
Nidetzky, Bernd [1 ,2 ]
机构：
[1] Austrian Ctr Ind Biotechnol, A-8010 Graz, Austria
[2] Graz Univ Technol, Inst Biotechnol & Biochem Engn, A-8010 Graz, Austria
[3] Graz Univ Technol, Inst Mol Biotechnol, A-8010 Graz, Austria
[4] Ingenza Ltd, Roslin EH25 9PP, Midlothian, Scotland
来源：
BIOTECHNOLOGY LETTERS | 2011年 / 33卷 / 03期
关键词：
D-Amino acid oxidase; Permeabilization; Pichia pastoris; Rhodotorula gracilis; Stability; TRIGONOPSIS-VARIABILIS; CATALYTIC-PROPERTIES; CEPHALOSPORIN-C; STABILIZATION; IMMOBILIZATION; MECHANISM; PROTEIN; ENZYME;
D O I：
10.1007/s10529-010-0456-9
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
By combining gene design and heterologous over-expression of Rhodotorula gracilis d-amino acid oxidase (RgDAO) in Pichia pastoris, enzyme production was enhanced by one order of magnitude compared to literature benchmarks, giving 350 kUnits/l of fed-batch bioreactor culture with a productivity of 3.1 kUnits/l h. P. pastoris cells permeabilized by freeze-drying and incubation in 2-propanol (10% v/v) produce a highly active (1.6 kUnits/g dry matter) and stable oxidase preparation. Critical bottlenecks in the development of an RgDAO catalyst for industrial applications have been eliminated.
引用
收藏
页码：557 / 563
页数：7
相关论文
共 26 条
  • [1] Stepwise engineering of a Pichia pastoris D-amino acid oxidase whole cell catalyst
    Abad, Sandra
    Nahalka, Jozef
    Bergler, Gabriele
    Arnold, S. Alison
    Speight, Robert
    Fotheringham, Ian
    Nidetzky, Bernd
    Glieder, Anton
    [J]. MICROBIAL CELL FACTORIES, 2010, 9
  • [2] Real-time PCR-based determination of gene copy numbers in Pichia pastoris
    Abad, Sandra
    Kitz, Kerstin
    Hoermann, Astrid
    Schreiner, Ulrike
    Hartner, Franz S.
    Glieder, Anton
    [J]. BIOTECHNOLOGY JOURNAL, 2010, 5 (04) : 413 - 420
  • [3] Deracemisation and stereoinversion of α-amino acids using D-amino acid oxidase and hydride reducing agents
    Beard, TM
    Turner, NJ
    [J]. CHEMICAL COMMUNICATIONS, 2002, (03) : 246 - 247
  • [4] Use of physicochemical tools to determine the choice of optimal enzyme:: Stabilization of D-amino acid oxidase
    Betancor, L
    Hidalgo, A
    Fernández-Lorente, G
    Mateo, C
    Rodríguez, V
    Fuentes, M
    López-Gallego, F
    Fernández-Lafuente, R
    Guisan, JM
    [J]. BIOTECHNOLOGY PROGRESS, 2003, 19 (03) : 784 - 788
  • [5] Trigonopsis variabilis D-amino acid oxidase:: Control of protein quality and opportunities for biocatalysis through production in Escherichia coli
    Dib, Iskandar
    Stanzer, Damir
    Nidetzky, Bernd
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (01) : 331 - 333
  • [6] Thermal inactivation of D-amino acid oxidase from Trigonopsis variabilis occurs via three parallel paths of irreversible denaturation
    Dib, Iskandar
    Slavica, Anita
    Riethorst, Waander
    Nidetzky, Bernd
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 2006, 94 (04) : 645 - 654
  • [7] The stabilizing effects of immobilization in D-amino acid oxidase from Trigonopsis variabilis
    Dib, Iskandar
    Nidetzky, Bernd
    [J]. BMC BIOTECHNOLOGY, 2008, 8 (1)
  • [8] Oxidation of amines by flavoproteins
    Fitzpatrick, Paul F.
    [J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2010, 493 (01) : 13 - 25
  • [9] Preparative deracemization of unnatural amino acids
    Fotheringham, I
    Archer, I
    Carr, R
    Speight, R
    Turner, NJ
    [J]. BIOCHEMICAL SOCIETY TRANSACTIONS, 2006, 34 : 287 - 290
  • [10] Hellwig S, 1999, BIOTECHNOL APPL BIOC, V30, P267
123