Skip to main content
SpringerLink
Log in

ATP-citrate lyase activity and carotenoid production in batch cultures of Phaffia rhodozyma under nitrogen-limited and nonlimited conditions

  • Applied Microbial and Cell Physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

ATP-citrate lyase (ACL) is the key cytoplasmic enzyme which supplies acetyl-CoA for fatty acids in oleaginous yeast. Although it has been suggested that fatty acid and carotenoid biosynthesis may have a common source of acetyl-CoA in Phaffia rhodozyma, the source for carotenoids is currently unknown. The purpose of this work was to analyze the development of ACL activity during batch cultures of P. rhodozyma under ammonium-limited and nonammonium-limited conditions and study its possible relationship with carotenoid synthesis. Every experiment showed carotenoid accumulation linked to an increasing ACL activity. Moreover, the ACL activity increased with dissolved oxygen (DO), i.e., ACL responded to DO in a similar way as carotenoid synthesis. Additionally, in the ammonium-limited culture, ACL activity increased upon ammonium depletion. However, the contribution to carotenoid accumulation in that case was negligible. This suggests that P. rhodozyma has developed two components of ACL, each one responsive to a different environmental stimulus, i.e., DO and ammonium depletion. The role of each component is still unknown; however, considering that the former responds to DO and the known role of carotenoids as antioxidants, it may be a provider of acetyl-CoA for carotenoid synthesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • An GH, Schuman DB, Johnson EA (1989) Isolation of Phaffia rhodozyma mutants with increased astaxanthin content. Appl Environ Microbiol 55:116–124

    CAS  Google Scholar 

  • Andlid T, Larsson C, Liljenberg C, Marison I, Gustafsson L (1995) Enthalpy content as a function of lipid accumulation in Rhodotorula glutinis. Appl Microbiol Biotechnol 42:818–825

    Article  CAS  Google Scholar 

  • Bhosale P (2004) Environmental and cultural stimulants in the production of carotenoids from microorganisms. Appl Microbiol Biotechnol 63(4):351–361

    Article  CAS  Google Scholar 

  • Bhosale P, Bernstein PS (2005) Microbial xanthophylls. Appl Microbiol Biotechnol 68(4):445–455

    Article  CAS  Google Scholar 

  • Boulton CA, Ratledge C (1981) Correlation of lipid accumulation in yeast with possession of ATP:citrate lyase. J Gen Microbiol 127:169–176

    CAS  Google Scholar 

  • Cannizzaro C, Christensen B, Nielsen J, Stockar UV (2004) Metabolic network analysis on Phaffia rhodozyma yeast using C-13-labeled glucose and gas chromatography-mass spectrometry. Metabolic Eng 6(4):340–351

    Article  CAS  Google Scholar 

  • Dufosse L (2006) Microbial production of food grade pigments. Food Technol Biotechnol 44(3):313–321

    CAS  Google Scholar 

  • Evans CT, Scragg AH, Ratledge C (1983) Regulation of citrate efflux from mitochondria of oleaginous and non-oleaginous yeast by adenine nucleotides. Eur J Biochem 132:609–615

    Article  CAS  Google Scholar 

  • Fell JW, Blatt GM (1999) Separation of strains of the yeast Xanthophyllomyces dendrorhous and Phaffia rhodozyma based on rDNA, IGS and ITS sequence analysis. J Ind Microbiol Biotechnol 23:677–681

    Article  CAS  Google Scholar 

  • Flores-Cotera LB, Martin R, Sánchez S (2001) Citrate, a possible precursor of astaxanthin in P. rhodozyma: influence of varying levels of ammonium, phosphate and citrate in a chemically defined medium. Appl Microbiol Biotechnol 55(3):341–347

    Article  CAS  Google Scholar 

  • Fraser PD, Miura Y, Misawa N (1997) In vitro characterization of astaxanthin biosynthetic enzymes. J Biol Chem 272:6128–6135

    Article  CAS  Google Scholar 

  • Gornall AG, Bardawill CJ, David MM (1949) Determination of serum proteins by means of the biuret reaction. J Biol Chem 177:751–766

    CAS  Google Scholar 

  • Gu WL, An GH, Johnson EA (1997) Ethanol increases carotenoid production in Phaffia rhodozyma. J Ind Microbiol Biotechnol 19:114–117

    Article  CAS  Google Scholar 

  • Johnson EA, Schroeder WA (1996) Biotechnology of astaxanthin production in Phaffia rhodozyma. In: Takeoka GR, Teranishi R, Williams PJ, Kobayashi A (eds) Biotechnology for improved foods and flavors. American Chemical Society, Washington DC, pp 39–50

    Chapter  Google Scholar 

  • Jones RP (1989) Biological principles for the effects of ethanol. Enz Microbial Technol 11(3):130–153

    Article  CAS  Google Scholar 

  • Kim W, Tabita FR (2006) Both subunits of ATP-citrate lyase from Chlorobium tepidum contribute to catalitic activity. J Bacteriol 188:6544–6552

    Article  CAS  Google Scholar 

  • Klionsky DT, Ohsumi Y (1999) Vacuolar import of proteins and organelles from the cytoplasm. Ann Rev Cell Dev Biol 15:1–32

    Article  CAS  Google Scholar 

  • Konz JO, King J, Cooney CL (1998) Effects of oxygen on recombinant protein expression. Biotechnol Prog 14:393–409

    Article  CAS  Google Scholar 

  • Lodato P, Alcaíno J, Barahona S, Niklitschek M, Carmona M, Wozniak A, Baeza M, Retamales P, Jimenez A, Cifuentes V (2007) Expression of the carotenoid biosynthesis genes in Xanthophyllomyces dendrorhous. Biol Res 40:73–84

    Article  CAS  Google Scholar 

  • Lopez S, Gancedo JM (1979) Effect of metabolic conditions on protein-turnover in yeast. Biochem J 178(3):769–776

    CAS  Google Scholar 

  • Meyer PS, du Preez JC (1994) Astaxanthin production by a Phaffia rhodozyma mutant on grape juice. World J Microbiol Biotechnol 10:178–183

    Article  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Nakano T, Kanmuri T, Sato M, Takeuchi M (1999) Effect of astaxanthin rich red yeast (Phaffia rhodozyma) on oxidative stress in rainbow trout. Biochim Biophys Acta 1426:119–125

    CAS  Google Scholar 

  • Ratledge CA, Wynn JP (2000) Understanding microbial obesity. SIM 50(4):181–185

    Google Scholar 

  • Ratledge C, Wynn JP (2002) The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. Adv Appl Microbiol 51:1–51

    Article  CAS  Google Scholar 

  • SAS Institute Inc (1989) Statistic guide for personal computers. Version 6.04. SAS Institute, Inc., NC, USA

    Google Scholar 

  • Schroeder WA, Johnson EA (1993) Antioxidant role of carotenoids in Phaffia rhodozyma. J Gen Microbiol 139:907–912

    CAS  Google Scholar 

  • Schroeder WA, Johnson EA (1995) Singlet oxygen and peroxyl radicals regulate carotenoid biosynthesis in Phaffia rhodozyma. J Biol Chem 270:18374–18379

    Article  CAS  Google Scholar 

  • Schuller HJ (2003) Transcriptional control of nonfermentative metabolism in the yeast Saccharomyces cerevisiae. Current Gen 43(3):139–160

    Google Scholar 

  • Sedmak JJ, Weerasinghe K, Jolly SO (1990) Extraction and quantitation of astaxanthin from Phaffia rhodozyma. Biotechnol Tech 4:107–112

    Article  CAS  Google Scholar 

  • Shashi K, Bachhawat AK, Joseph R (1990) ATP:citrate lyase of Rhodotorula gracilis: purification and properties. Biochim Biophys Acta 1033:23–30

    CAS  Google Scholar 

  • Sreré PA (1959) The citrate cleavage enzyme. I Distribution and purification. J Biological Chem 234(10):2544–2547

    Google Scholar 

  • Vries SD, Marres CAM (1987) The mitochondrial respiratory chain of yeast. Structure and biosynthesis and the role in cellular metabolism. Biochim Biophys Acta 895:205–239

    Google Scholar 

  • Vustin MM, Belykh EN, Kishilova SA (2004) Relationship between astaxanthin production and the intensity of anabolic processes in the yeast Phaffia rhodozyma. Microbiology 73(6):643–649

    Article  CAS  Google Scholar 

  • Weatherburn MW (1967) Phenol-hypochlorite reaction for determination of ammonia. Anal Chem 39:971–973

    Article  CAS  Google Scholar 

  • Yamane Y, Higashida K, Nakashimada Y, Kakizono T, Nishio N (1997a) Astaxanthin production by Phaffia rhodozyma enhanced in fed-batch culture with glucose and ethanol feeding. Biotechnol Lett 19:1109–1111

    Article  CAS  Google Scholar 

  • Yamane Y, Higashida K, Nakashimada Y, Kakizono T, Nishio N (1997b) Influence of oxygen and glucose on primary metabolism and astaxanthin production by Phaffia rhodozyma in batch and fed-batch cultures: kinetic and stoichiometric analysis. Appl Environ Microbiol 63:4471–4478

    CAS  Google Scholar 

Download references

Acknowledgements

The authors thank to Elvira Ríos-Leal for assistance in ethanol analyses and Fernando Maldonado-García and Jesus Vega-Estrada for and their expert technical assistance. The research was funded by the Department of Biotechnology and Bioengineering, Cinvestav.

Author information

Authors and Affiliations

  1. Department of Biotechnology and Bioengineering, Cinvestav-IPN, Av. Instituto Politécnico Nacional 2508, Col. San Pedro Zacatenco, México, 07360, DF, México

    Cipriano Chávez-Cabrera, Zoila R. Flores-Bustamante, Rodolfo Marsch, María del Carmen Montes & Luis Bernardo Flores-Cotera

  2. Department of Biotechnology, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, DF, 04510, México

    Sergio Sánchez

  3. Department of Microbiology, Escuela Nacional de Ciencias Biológicas del IPN, México, DF, 11340, México

    Juan Carlos Cancino-Díaz

Authors
  1. Cipriano Chávez-Cabrera
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zoila R. Flores-Bustamante
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rodolfo Marsch
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. María del Carmen Montes
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sergio Sánchez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juan Carlos Cancino-Díaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luis Bernardo Flores-Cotera
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Luis Bernardo Flores-Cotera.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chávez-Cabrera, C., Flores-Bustamante, Z.R., Marsch, R. et al. ATP-citrate lyase activity and carotenoid production in batch cultures of Phaffia rhodozyma under nitrogen-limited and nonlimited conditions. Appl Microbiol Biotechnol 85, 1953–1960 (2010). https://doi.org/10.1007/s00253-009-2271-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-009-2271-6

Keywords

Search

Navigation