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Papers by Stephan Mauersberger
Appl Microbiol Biotechnol, 1992
In Candida maltosa and other alkene-utilizing yeasts a membrane-bound fatty alcohol oxidase (FAOD... more In Candida maltosa and other alkene-utilizing yeasts a membrane-bound fatty alcohol oxidase (FAOD) is induced by growth on n-alkenes. The oxidation of 1-alkanols to the corresponding aldehydes is accompanied by the stoichiometric consumption of 1 mol O 2 ...
GBM Annual Fall meeting Halle 2002, 2002
Microbiology Monographs, 2013
Microbiology Monographs, 2013
Since the first studies in 1980s, P450 has been shown to be induced during growth of Y. lipolytic... more Since the first studies in 1980s, P450 has been shown to be induced during growth of Y. lipolytica on alkanes, fatty alcohols and fatty acids and evidenced to be involved in terminal hydroxylation of alkanes and ω-hydroxylation of fatty acids without differentiation between P450 isoforms. The alkane-hydroxylating P450 exhibit an extraordinary high in vivo activity with turnover numbers up to 3,000/min.
Applied Microbiology and Biotechnology, Apr 20, 2007
The yeast Yarrowia lipolytica is able to secrete high amounts of several organic acids under cond... more The yeast Yarrowia lipolytica is able to secrete high amounts of several organic acids under conditions of growth limitation and carbon source excess. Here we report the production of citric acid (CA) in a fed-batch cultivation process on sucrose using the recombinant Y. lipolytica strain H222-S4(p67ICL1) T5, harbouring the invertase encoding ScSUC2 gene of Saccharomyces cerevisiae under the inducible XPR2 promoter control and multiple ICL1
Yeast, 2005
The novel LTR retrotransposon Tyl6 was detected in the genome of the dimorphic fungus Yarrowia li... more The novel LTR retrotransposon Tyl6 was detected in the genome of the dimorphic fungus Yarrowia lipolytica. Sequence analysis revealed that this element is related to the well-known Ty3 element of Saccharomyces cerevisiae and, especially, to the recently described Tse3 retrotransposon of Saccharomyces exiguus and to the del1-like plant retrotransposons. Tyl6 is 5108 bp long, is flanked by two identical long terminal repeats (LTR), each of 276 bp, and its ORFs are separated by a −1 frameshift. Both ORFs are intact and deduced translation products display a significant similarity with those of previously described Ty3/gypsy retrotransposons. Distribution of Tyl6 among Y. lipolytica strains of different origins was also analysed. A single copy of the novel retrotransposon is present in some commonly used laboratory strains, which are derivatives of the wild-type isolate YB423-12, whereas other strains of independent origin are devoid of Ty16. No solo LTR of Tyl6 was detected in the analysed strains. The complete sequence of Tyl6 was submitted to the EMBL nucleotide sequence database under Accession No. AJ746250.
Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry, 2007
Using recombinant microorganisms S. cerevisiae GRF18/YEp 5117 α , expressing bovine adrenocortica... more Using recombinant microorganisms S. cerevisiae GRF18/YEp 5117 α , expressing bovine adrenocortical cytochrome P450cl7, we have studied the effect of various modifiers of steroid biosynthesis on the relationship between reactions of the 17 α -hydroxylation and 20 α -reduction of progesterone. Dexamethasone and metyrapone had no effect on the reaction of progesterone 17 α -hydroxylation and 20 α -reduction of 17 α -hydroxyprogesterone. Mifepriston and danazol did not covalently modify amino acid residues of the cytochrome P450cl7 or its heme group under the conditions of progesterone biotransformation by recombinant yeasts. Ketokonazole, mifepriston and danazol were found to be low-affinity competitive inhibitors, but the 20-dihydroderivatives of progesterone were mixed type inhibitors of the cytochrome P450cl7. All modifiers used did not affect the functional properties of the yeast analog of 20 α -hydroxysteroid dehydrogenase. Based on the effect on catalytic parameters of the cytochrome P450cl7, the all modifiers used can be arranged in the following order: 20 β -dihydroprogesterone (maximal effect) > mifepriston = ketokonazole > 20 α -dihydroprogesterone > danazol > dexamethasone, metyrapone (without effect).
Mikrobiologiia
The content of cytochrome P-450 was studied in the cells of alkane oxidizing yeasts Candida guill... more The content of cytochrome P-450 was studied in the cells of alkane oxidizing yeasts Candida guilliermondii, C. tropicalis and C. lipolytica. The cells of all the studied yeast strains growing on hexadecane were found to contain cytochrome P-450. The cytochrome was not detected when the yeast strains grew on glucose. The concentration of cytochrome P-450 remained constant at the exponential growth phase, but decreased at the beginning of the stationary growth phase. Cytochrome P-450 was shown to be synthesized de novo in the course of physiological adaptation of the cells to hexadecane.
European Journal of Cell Biology
ABSTRACT
To induce the overproduction of citric and isocitric acids at different proportions, as well as a... more To induce the overproduction of citric and isocitric acids at different proportions, as well as a-ketoglutaric acid, in Y. lipolytica by changing the cultivation conditions.
Non-Conventional Yeasts in Genetics, Biochemistry and Biotechnology, 2003
Non-Conventional Yeasts in Genetics, Biochemistry and Biotechnology, 2003
Nonconventional Yeasts in Biotechnology, 1996
Biochemistry. Biokhimii͡a, 2002
The cDNA encoding cytochrome P-45017alpha from bovine adrenal cortex was expressed in Saccharomyc... more The cDNA encoding cytochrome P-45017alpha from bovine adrenal cortex was expressed in Saccharomyces cerevisiae under the control of the galactose-inducible GAL10 promoter. Carbon monoxide difference spectra of the galactose-induced yeast cells showed expression of about 240 nmol of P-45017alpha per liter of the culture. Binding of progesterone to the cytochrome P-45017alpha was clearly detectable already with intact yeast cells as judged by the formation of type I substrate difference spectra. Yeast cells grown on minimal medium containing galactose actively converted progesterone to 17alpha-hydroxyprogesterone, this indicating the functional integrity of the heterologously expressed P-45017alpha and its efficient coupling with the constitutive NADPH-cytochrome P-450 reductase. More than 80% of the metabolite produced was secreted into the culture medium. Cultivation in a rich non-selective medium resulted in the formation of an additional product, which was identified by mass spect...
Microbial Cell Factories, 2012
Background: Yarrowia lipolytica efficiently metabolizes and assimilates hydrophobic compounds suc... more Background: Yarrowia lipolytica efficiently metabolizes and assimilates hydrophobic compounds such as n-alkanes and fatty acids. Efficient substrate uptake is enabled by naturally secreted emulsifiers and a modified cell surface hydrophobicity and protrusions formed by this yeast. We were examining the potential of recombinant Y. lipolytica as a biocatalyst for the oxidation of hardly soluble hydrophobic steroids. Furthermore, two-liquid biphasic culture systems were evaluated to increase substrate availability. While cells, together with water soluble nutrients, are maintained in the aqueous phase, substrates and most of the products are contained in a second water-immiscible organic solvent phase.
Zeitschrift für allgemeine Mikrobiologie, 1983
The utilization of n-alkanes is connected with extensive modifications of the yeast cell, especia... more The utilization of n-alkanes is connected with extensive modifications of the yeast cell, especially of the cytochrome P-450-containing membrane. Beside the cytochrome P-450 the NADPH cytochrome P-450 reductasa, the cytochrome b6, long-chain alcohol and long-chain aldehyde dehydrogenases are induced. The activity of the alkane-hydroxylating enzyme system grows more than the concentration of its terminal oxidase. The induction of the cytochrome P-460 is inhibited by cyclcheximide. A low concentration of oxygen in the culture medium amplifies the induction both of the alkane-hydroxylating enzyme system and of catalase and cytochrome oxidase, which arelocalized in the peroxisomes and mitochondria, respectively.
Zeitschrift für allgemeine Mikrobiologie, 1981
Fungal Genetics and Biology, 2007
In order to get deeper insights into oxidative degradation of the hydrophobic substrates (HS) tri... more In order to get deeper insights into oxidative degradation of the hydrophobic substrates (HS) triglycerides and alkanes by yeasts, tagged mutants affected in these pathways were generated by random insertion of a mutagenesis cassette MTC into the genome of Yarrowia lipolytica. About 9.600 Ura+ transformants were screened in plate tests for utilization of alkanes (C10, C16), oleic acid and tributyrin. HS degradation mutants were recovered as unable to grow on alkane or on intermediates of the pathway (AlkA-AlkE phenotype classes). To identify the disrupted genes, insertion points of the MTC were sequenced using convergent and divergent PCR. Sequence analysis evidenced both known and new genes required for HS utilization, e.g. for AlkD/E mutants MTC insertion had occurred in genes of thioredoxin reductase, peroxines PEX14 and PEX20, succinate-fumarate carrier SFC1, and isocitrate lyase ICL1. Several mutants were affected in alkane utilization depending on chain length. Mutant Z110 (AlkAb: C10- C16+) was shown to be disrupted for ANT1 encoding a peroxisomal membrane localized adenine nucleotide transporter protein, providing ATP for the activation of short-chain fatty acids by acyl-CoA synthetase II in peroxisomes. Mutants N046 and B095 (AlkAc: C10+ C16-) were disrupted for the ABC transporter encoded by ABC1 gene, thus providing first evidence for its participation in chain length dependent alkane transport processes.
Appl Microbiol Biotechnol, 1992
In Candida maltosa and other alkene-utilizing yeasts a membrane-bound fatty alcohol oxidase (FAOD... more In Candida maltosa and other alkene-utilizing yeasts a membrane-bound fatty alcohol oxidase (FAOD) is induced by growth on n-alkenes. The oxidation of 1-alkanols to the corresponding aldehydes is accompanied by the stoichiometric consumption of 1 mol O 2 ...
GBM Annual Fall meeting Halle 2002, 2002
Microbiology Monographs, 2013
Microbiology Monographs, 2013
Since the first studies in 1980s, P450 has been shown to be induced during growth of Y. lipolytic... more Since the first studies in 1980s, P450 has been shown to be induced during growth of Y. lipolytica on alkanes, fatty alcohols and fatty acids and evidenced to be involved in terminal hydroxylation of alkanes and ω-hydroxylation of fatty acids without differentiation between P450 isoforms. The alkane-hydroxylating P450 exhibit an extraordinary high in vivo activity with turnover numbers up to 3,000/min.
Applied Microbiology and Biotechnology, Apr 20, 2007
The yeast Yarrowia lipolytica is able to secrete high amounts of several organic acids under cond... more The yeast Yarrowia lipolytica is able to secrete high amounts of several organic acids under conditions of growth limitation and carbon source excess. Here we report the production of citric acid (CA) in a fed-batch cultivation process on sucrose using the recombinant Y. lipolytica strain H222-S4(p67ICL1) T5, harbouring the invertase encoding ScSUC2 gene of Saccharomyces cerevisiae under the inducible XPR2 promoter control and multiple ICL1
Yeast, 2005
The novel LTR retrotransposon Tyl6 was detected in the genome of the dimorphic fungus Yarrowia li... more The novel LTR retrotransposon Tyl6 was detected in the genome of the dimorphic fungus Yarrowia lipolytica. Sequence analysis revealed that this element is related to the well-known Ty3 element of Saccharomyces cerevisiae and, especially, to the recently described Tse3 retrotransposon of Saccharomyces exiguus and to the del1-like plant retrotransposons. Tyl6 is 5108 bp long, is flanked by two identical long terminal repeats (LTR), each of 276 bp, and its ORFs are separated by a −1 frameshift. Both ORFs are intact and deduced translation products display a significant similarity with those of previously described Ty3/gypsy retrotransposons. Distribution of Tyl6 among Y. lipolytica strains of different origins was also analysed. A single copy of the novel retrotransposon is present in some commonly used laboratory strains, which are derivatives of the wild-type isolate YB423-12, whereas other strains of independent origin are devoid of Ty16. No solo LTR of Tyl6 was detected in the analysed strains. The complete sequence of Tyl6 was submitted to the EMBL nucleotide sequence database under Accession No. AJ746250.
Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry, 2007
Using recombinant microorganisms S. cerevisiae GRF18/YEp 5117 α , expressing bovine adrenocortica... more Using recombinant microorganisms S. cerevisiae GRF18/YEp 5117 α , expressing bovine adrenocortical cytochrome P450cl7, we have studied the effect of various modifiers of steroid biosynthesis on the relationship between reactions of the 17 α -hydroxylation and 20 α -reduction of progesterone. Dexamethasone and metyrapone had no effect on the reaction of progesterone 17 α -hydroxylation and 20 α -reduction of 17 α -hydroxyprogesterone. Mifepriston and danazol did not covalently modify amino acid residues of the cytochrome P450cl7 or its heme group under the conditions of progesterone biotransformation by recombinant yeasts. Ketokonazole, mifepriston and danazol were found to be low-affinity competitive inhibitors, but the 20-dihydroderivatives of progesterone were mixed type inhibitors of the cytochrome P450cl7. All modifiers used did not affect the functional properties of the yeast analog of 20 α -hydroxysteroid dehydrogenase. Based on the effect on catalytic parameters of the cytochrome P450cl7, the all modifiers used can be arranged in the following order: 20 β -dihydroprogesterone (maximal effect) > mifepriston = ketokonazole > 20 α -dihydroprogesterone > danazol > dexamethasone, metyrapone (without effect).
Mikrobiologiia
The content of cytochrome P-450 was studied in the cells of alkane oxidizing yeasts Candida guill... more The content of cytochrome P-450 was studied in the cells of alkane oxidizing yeasts Candida guilliermondii, C. tropicalis and C. lipolytica. The cells of all the studied yeast strains growing on hexadecane were found to contain cytochrome P-450. The cytochrome was not detected when the yeast strains grew on glucose. The concentration of cytochrome P-450 remained constant at the exponential growth phase, but decreased at the beginning of the stationary growth phase. Cytochrome P-450 was shown to be synthesized de novo in the course of physiological adaptation of the cells to hexadecane.
European Journal of Cell Biology
ABSTRACT
To induce the overproduction of citric and isocitric acids at different proportions, as well as a... more To induce the overproduction of citric and isocitric acids at different proportions, as well as a-ketoglutaric acid, in Y. lipolytica by changing the cultivation conditions.
Non-Conventional Yeasts in Genetics, Biochemistry and Biotechnology, 2003
Non-Conventional Yeasts in Genetics, Biochemistry and Biotechnology, 2003
Nonconventional Yeasts in Biotechnology, 1996
Biochemistry. Biokhimii͡a, 2002
The cDNA encoding cytochrome P-45017alpha from bovine adrenal cortex was expressed in Saccharomyc... more The cDNA encoding cytochrome P-45017alpha from bovine adrenal cortex was expressed in Saccharomyces cerevisiae under the control of the galactose-inducible GAL10 promoter. Carbon monoxide difference spectra of the galactose-induced yeast cells showed expression of about 240 nmol of P-45017alpha per liter of the culture. Binding of progesterone to the cytochrome P-45017alpha was clearly detectable already with intact yeast cells as judged by the formation of type I substrate difference spectra. Yeast cells grown on minimal medium containing galactose actively converted progesterone to 17alpha-hydroxyprogesterone, this indicating the functional integrity of the heterologously expressed P-45017alpha and its efficient coupling with the constitutive NADPH-cytochrome P-450 reductase. More than 80% of the metabolite produced was secreted into the culture medium. Cultivation in a rich non-selective medium resulted in the formation of an additional product, which was identified by mass spect...
Microbial Cell Factories, 2012
Background: Yarrowia lipolytica efficiently metabolizes and assimilates hydrophobic compounds suc... more Background: Yarrowia lipolytica efficiently metabolizes and assimilates hydrophobic compounds such as n-alkanes and fatty acids. Efficient substrate uptake is enabled by naturally secreted emulsifiers and a modified cell surface hydrophobicity and protrusions formed by this yeast. We were examining the potential of recombinant Y. lipolytica as a biocatalyst for the oxidation of hardly soluble hydrophobic steroids. Furthermore, two-liquid biphasic culture systems were evaluated to increase substrate availability. While cells, together with water soluble nutrients, are maintained in the aqueous phase, substrates and most of the products are contained in a second water-immiscible organic solvent phase.
Zeitschrift für allgemeine Mikrobiologie, 1983
The utilization of n-alkanes is connected with extensive modifications of the yeast cell, especia... more The utilization of n-alkanes is connected with extensive modifications of the yeast cell, especially of the cytochrome P-450-containing membrane. Beside the cytochrome P-450 the NADPH cytochrome P-450 reductasa, the cytochrome b6, long-chain alcohol and long-chain aldehyde dehydrogenases are induced. The activity of the alkane-hydroxylating enzyme system grows more than the concentration of its terminal oxidase. The induction of the cytochrome P-460 is inhibited by cyclcheximide. A low concentration of oxygen in the culture medium amplifies the induction both of the alkane-hydroxylating enzyme system and of catalase and cytochrome oxidase, which arelocalized in the peroxisomes and mitochondria, respectively.
Zeitschrift für allgemeine Mikrobiologie, 1981
Fungal Genetics and Biology, 2007
In order to get deeper insights into oxidative degradation of the hydrophobic substrates (HS) tri... more In order to get deeper insights into oxidative degradation of the hydrophobic substrates (HS) triglycerides and alkanes by yeasts, tagged mutants affected in these pathways were generated by random insertion of a mutagenesis cassette MTC into the genome of Yarrowia lipolytica. About 9.600 Ura+ transformants were screened in plate tests for utilization of alkanes (C10, C16), oleic acid and tributyrin. HS degradation mutants were recovered as unable to grow on alkane or on intermediates of the pathway (AlkA-AlkE phenotype classes). To identify the disrupted genes, insertion points of the MTC were sequenced using convergent and divergent PCR. Sequence analysis evidenced both known and new genes required for HS utilization, e.g. for AlkD/E mutants MTC insertion had occurred in genes of thioredoxin reductase, peroxines PEX14 and PEX20, succinate-fumarate carrier SFC1, and isocitrate lyase ICL1. Several mutants were affected in alkane utilization depending on chain length. Mutant Z110 (AlkAb: C10- C16+) was shown to be disrupted for ANT1 encoding a peroxisomal membrane localized adenine nucleotide transporter protein, providing ATP for the activation of short-chain fatty acids by acyl-CoA synthetase II in peroxisomes. Mutants N046 and B095 (AlkAc: C10+ C16-) were disrupted for the ABC transporter encoded by ABC1 gene, thus providing first evidence for its participation in chain length dependent alkane transport processes.