Comparative proteome analysis of Hansenula polymorpha DL1 and A16

Abstract

Proteomic responses of methylotrophic yeasts (Hansenula polymorpha DL1 and A16) to growth medium tuning by carbon source shift (glycerol-->methanol) were monitored and analyzed by two-dimensional gel electrophoresis. Through comparative analyses of two-dimensional gels, intracellular yeast proteins with complex expression patterns were systematically sorted into: (1) proteins that are commonly expressed with comparable high abundance in both strains; (2) strain-specific proteins that are expressed at high level only in a particular strain; (3) strain-specific and methanol-induced proteins that are expressed only in the presence of methanol; and (4) strain-specific and constitutively-expressed proteins that are expressed consistently irrespective of carbon source shift without extreme change in expression level. Among the DL1-specific proteins belonging to group four, the four proteins showing the highest expression levels in the course of the fermentation process were identified as: glucose-6-phosphate dehydrogenase, isocitrate lyase, succinyl-CoA synthetase, and glycerol-3-phosphate dehydrogenase. From these results, it is suggested that DL1 has distinct metabolic characteristics including enhanced metabolic activities both in glycerol uptake and the glyoxylate bypass cycle, as compared to A16. This is likely to explain why the DL1 strain shows a significantly higher rate of glycerol and methanol consumption during the fermentation process. Our systematic approach to the analysis of proteomic responses and the detailed analysis results reported here will be useful to better understand the global physiology of H. polymorpha, as proteome databases for various methylotrophic yeasts are established.

Yeast Proteome Analysis

“…Proteins specific for the DL1 strain, but constantly expressed under both carbon sources included glucose-6-phosphate dehydrogenase, isocitrate lyase, scuccinyl-CoA synthetase and glycerol-3-phsophae dehydrogenase. These findings correlated with the higher rate of glycerol and methanol consumption in DL1 strain (Kim et al, 2004 ).…”

“…A comparative proteome analysis of the methylotrophic Hansenula polymorpha strains DL1 and A16 was performed to monitor the changes in metabolism when shifting from glycerol to methanol as a carbon source (Kim et al, 2004 ). The facultative methylotroph H. polymorpha has been shown to be a good host system for the expression of heterologous proteins, and is in use for industrial production of proteins.…”

Escherichia coli malate dehydrogenase, a novel solubility enhancer for heterologous proteins synthesized in Escherichia coli

“…After sonication, the cell debris was removed by centrifugation at 16,000g for 60 min (48C). A 2-dimensional gel electrophoresis was performed as described previously Kim et al (2004).…”

Genome sequence and analysis of methylotrophic yeast Hansenula polymorpha DL1

“…The H. polymorpha species complex in fact includes several phylogenetically distinct strains [21] now reclassified as Ogataea polymorpha and Ogataea parapolymorpha [22,23]. A genome sequencing project for strain H. polymorpha CBS4732 that resulted in assembly of about 90% of the genome, including the vast majority of encoded proteins [24], appears extremely useful for comparative genomic and proteomic studies [21,25], identification of various transcription responses [26] and studies of mechanisms of strain adaptation to growth on methanol [27]. …”