Lex de Boer - Academia.edu (original) (raw)

Papers by Lex de Boer

Abstract. Cytoplasmic membrane vesicles were prepared by a lysozyme-salt treatment from Arthrobac... more Abstract. Cytoplasmic membrane vesicles were prepared by a lysozyme-salt treatment from Arthrobacter P] grown on methylamine asthe carbon and energy source. In the presence of an ascorbate-phenazine m thosulphate electron donor system, these vesicles accumulated methylamine in unmodi-fied form by an inducible transport system. This system has a high affinity for methylamine (Kapp = 20- 25 pM). The effect of the ionophores valinomycin and nigericin combined with membrane potential (A0) and pH-gradient (ApH) measure-ments demonstrated that methylamine uptake is electrogenic and driven by the A 0. Optimal activity is observed at pH 6.5 and 30 ~ Methylamine uptake was not affected by the presence of ammonium ions but was inhibited by the primary amines ethylamine (competitively), propylamine, butylamine and benzylamine. In addition, formaldehyde and acetate, at a concentration of1 mM, inhibited methylamine uptake almost completely. These compounds were shown to be non-com-petitive inhib...

Archives of Microbiology, 1984

Archives of Microbiology, 1982

Applied Microbiology and Biotechnology, 1990

Antonie van Leeuwenhoek, 1989

The metabolism of trimethylamine (TMA) and dimethylamine (DMA) in Arthrobacter P1 involved the en... more The metabolism of trimethylamine (TMA) and dimethylamine (DMA) in Arthrobacter P1 involved the enzymes TMA monooxygenase and trimethylamine-N-oxide (TMA-NO) demethylase, and DMA monooxygenase, respectively. The methylamine and formaldehyde produced were further metabolized via a primary amine oxidase and the ribulose monophosphate (RUMP) cycle. The amine oxidase showed activity with various aliphatic primary amines and benzylamine. The organism was able to use methylamine, ethylamine and propylamine as carbonand nitrogen sources for growth. Butylamine and benzylamine only functioned as nitrogen sources. Growth on glucose with ethylamine, propylamine, butylamine and benzylamine resulted in accumulation of the respective aldehydes. In case of ethylamine and propylamine this was due to repression by glucose of the synthesis of the aldehyde dehydrogenase(s) required for their further metabolism. Growth on glucose/methylamine did not result in repression of the RuMP cycle enzyme hexulose-6-phosphate synthase (HPS). High levels of this enzyme were present in the cells and as a result formaldehyde did not accumulate. Ammonia assimilation in Arthrobacter P1 involved NADP-dependent glutamate dehydrogenase (GDH), NAD-dependent alanine dehydrogenase (ADH) and glutamine synthetase (GS) as key enzymes. In batch cultures both GDH and GS displayed highest levels during growth on acetate with methylamine as the nitrogen source. A further increase in the levels of GS, but not GDH, was observed under ammonia-limited growth conditions in continuous cultures with acetate or glucose as carbon sources.

Antonie van Leeuwenhoek, 1985

Archives of Microbiology, 1990

The regulation of methanol metabolism in Nocardia sp. 239 was investigated. Growth on mixtures of... more The regulation of methanol metabolism in Nocardia sp. 239 was investigated. Growth on mixtures of glucose or acetate plus methanol in batch cultures resulted in simultaneous utilization of the substrates. The presence of glucose, but not of acetate, repressed synthesis of the ribulose monophosphate (RUMP) cycle enzymes hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), and methanol was used as an energy source only. Comparable results were obtained following addition of formaldehyde (fedbatch system) to a culture growing on glucose. The synthesis of the methanol dissimilatory and assimilatory enzymes in Nocardia sp. 239 thus appears to be controlled differently. Methanol and/or formaldehyde induce the synthesis of these enzymes, but under carbon-excess conditions their inducing effect on HPS and HPI synthesis is completely overruled by glucose, or metabolites derived from it. Repression of the synthesis of these RuMP cycle enzymes was of minor importance under carbon-and energy-limiting conditions in chemostat cultures. Addition of a pulse of glucose to a formaldehyde-limited (2.5 mmol 1-a h-1) fed-batch culture resulted in a decrease in the levels of several enzymes of methanol metabolism (including HPI), whereas the HPS levels remained relatively constant. Increasing HPS/HPI activity ratios were also observed with increasing growth rates in formaldehyde-limited chemostat cultures. The data indicate that additional mechanisms, the identity of which remains to be elucidated, are involved in controlling the levels of these Ca-specific enzymes in Nocardia sp. 239.

Applied microbiology and biotechnology, 2008

Two extracellular enzymes (MsP1 and MsP2) capable of efficient beta-carotene degradation were pur... more Two extracellular enzymes (MsP1 and MsP2) capable of efficient beta-carotene degradation were purified from culture supernatants of the basidiomycete Marasmius scorodonius (garlic mushroom). Under native conditions, the enzymes exhibited molecular masses of approximately 150 and approximately 120 kDa, respectively. SDS-PAGE and mass spectrometric data suggested a composition of two identical subunits for both enzymes. Biochemical characterisation of the purified proteins showed isoelectric points of 3.7 and 3.5, and the presence of heme groups in the active enzymes. Partial amino acid sequences were derived from N-terminal Edman degradation and from mass spectrometric ab initio sequencing of internal peptides. cDNAs of 1,604 to 1,923 bp, containing open reading frames (ORF) of 508 to 513 amino acids, respectively, were cloned from a cDNA library of M. scorodonius. These data suggest glycosylation degrees of approximately 23% for MsP1 and 8% for MsP2. Databank homology searches revea...

Het doel van dit project was de realisatie van een proof-of-principle keten voor de productie van... more Het doel van dit project was de realisatie van een proof-of-principle keten voor de productie van astaxanthine als oleoresin uit de alg Haematococcus pluvialis in fotobioreactoren in Nederlandse kassen. Astaxanthine is een sterk antioxidant dat bij kan dragen aan een gezonde voeding voor consumenten. Astaxanthine kan in Nederlandse kassen duurzaam worden geproduceerd (kassen als zonnecollector, gebruik afval CO 2 uit industrie, reinigen afvalwater). Binnen dit project is onderzoek verricht naar de selectie van natuurlijke algenstammen op kasniveau, klassieke stamverbetering op labniveau (uitgangsmateriaal), procesverbetering in alle fases van de productie (voorkweek, opkweek, groene fase, rode fase) door optimalisatie teeltfactoren en hygiënisatie (teelt en productie) en het maken van een proof-of-principle eindproduct (eindformulering) volgens marktspecificatie (markt en economie). Met het project is kennis vergaard om een nieuw verdienmodel voor de productie van een hoogwaardige stof (astaxanthine) in de tuinbouw te realiseren. Er is een proof-of-principle van een economisch rendabele productieketen van uitgangsmateriaal over productie tot product eindformulering volgens specificaties van de markt aangetoond voor de productie van astaxanthine uit Haematococcus pluvialis.

Abstract. Cytoplasmic membrane vesicles were prepared by a lysozyme-salt treatment from Arthrobac... more Abstract. Cytoplasmic membrane vesicles were prepared by a lysozyme-salt treatment from Arthrobacter P] grown on methylamine asthe carbon and energy source. In the presence of an ascorbate-phenazine m thosulphate electron donor system, these vesicles accumulated methylamine in unmodi-fied form by an inducible transport system. This system has a high affinity for methylamine (Kapp = 20- 25 pM). The effect of the ionophores valinomycin and nigericin combined with membrane potential (A0) and pH-gradient (ApH) measure-ments demonstrated that methylamine uptake is electrogenic and driven by the A 0. Optimal activity is observed at pH 6.5 and 30 ~ Methylamine uptake was not affected by the presence of ammonium ions but was inhibited by the primary amines ethylamine (competitively), propylamine, butylamine and benzylamine. In addition, formaldehyde and acetate, at a concentration of1 mM, inhibited methylamine uptake almost completely. These compounds were shown to be non-com-petitive inhib...

Archives of Microbiology, 1984

Archives of Microbiology, 1982

Applied Microbiology and Biotechnology, 1990

Antonie van Leeuwenhoek, 1989

The metabolism of trimethylamine (TMA) and dimethylamine (DMA) in Arthrobacter P1 involved the en... more The metabolism of trimethylamine (TMA) and dimethylamine (DMA) in Arthrobacter P1 involved the enzymes TMA monooxygenase and trimethylamine-N-oxide (TMA-NO) demethylase, and DMA monooxygenase, respectively. The methylamine and formaldehyde produced were further metabolized via a primary amine oxidase and the ribulose monophosphate (RUMP) cycle. The amine oxidase showed activity with various aliphatic primary amines and benzylamine. The organism was able to use methylamine, ethylamine and propylamine as carbonand nitrogen sources for growth. Butylamine and benzylamine only functioned as nitrogen sources. Growth on glucose with ethylamine, propylamine, butylamine and benzylamine resulted in accumulation of the respective aldehydes. In case of ethylamine and propylamine this was due to repression by glucose of the synthesis of the aldehyde dehydrogenase(s) required for their further metabolism. Growth on glucose/methylamine did not result in repression of the RuMP cycle enzyme hexulose-6-phosphate synthase (HPS). High levels of this enzyme were present in the cells and as a result formaldehyde did not accumulate. Ammonia assimilation in Arthrobacter P1 involved NADP-dependent glutamate dehydrogenase (GDH), NAD-dependent alanine dehydrogenase (ADH) and glutamine synthetase (GS) as key enzymes. In batch cultures both GDH and GS displayed highest levels during growth on acetate with methylamine as the nitrogen source. A further increase in the levels of GS, but not GDH, was observed under ammonia-limited growth conditions in continuous cultures with acetate or glucose as carbon sources.

Antonie van Leeuwenhoek, 1985

Archives of Microbiology, 1990

The regulation of methanol metabolism in Nocardia sp. 239 was investigated. Growth on mixtures of... more The regulation of methanol metabolism in Nocardia sp. 239 was investigated. Growth on mixtures of glucose or acetate plus methanol in batch cultures resulted in simultaneous utilization of the substrates. The presence of glucose, but not of acetate, repressed synthesis of the ribulose monophosphate (RUMP) cycle enzymes hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), and methanol was used as an energy source only. Comparable results were obtained following addition of formaldehyde (fedbatch system) to a culture growing on glucose. The synthesis of the methanol dissimilatory and assimilatory enzymes in Nocardia sp. 239 thus appears to be controlled differently. Methanol and/or formaldehyde induce the synthesis of these enzymes, but under carbon-excess conditions their inducing effect on HPS and HPI synthesis is completely overruled by glucose, or metabolites derived from it. Repression of the synthesis of these RuMP cycle enzymes was of minor importance under carbon-and energy-limiting conditions in chemostat cultures. Addition of a pulse of glucose to a formaldehyde-limited (2.5 mmol 1-a h-1) fed-batch culture resulted in a decrease in the levels of several enzymes of methanol metabolism (including HPI), whereas the HPS levels remained relatively constant. Increasing HPS/HPI activity ratios were also observed with increasing growth rates in formaldehyde-limited chemostat cultures. The data indicate that additional mechanisms, the identity of which remains to be elucidated, are involved in controlling the levels of these Ca-specific enzymes in Nocardia sp. 239.

Applied microbiology and biotechnology, 2008

Two extracellular enzymes (MsP1 and MsP2) capable of efficient beta-carotene degradation were pur... more Two extracellular enzymes (MsP1 and MsP2) capable of efficient beta-carotene degradation were purified from culture supernatants of the basidiomycete Marasmius scorodonius (garlic mushroom). Under native conditions, the enzymes exhibited molecular masses of approximately 150 and approximately 120 kDa, respectively. SDS-PAGE and mass spectrometric data suggested a composition of two identical subunits for both enzymes. Biochemical characterisation of the purified proteins showed isoelectric points of 3.7 and 3.5, and the presence of heme groups in the active enzymes. Partial amino acid sequences were derived from N-terminal Edman degradation and from mass spectrometric ab initio sequencing of internal peptides. cDNAs of 1,604 to 1,923 bp, containing open reading frames (ORF) of 508 to 513 amino acids, respectively, were cloned from a cDNA library of M. scorodonius. These data suggest glycosylation degrees of approximately 23% for MsP1 and 8% for MsP2. Databank homology searches revea...

Het doel van dit project was de realisatie van een proof-of-principle keten voor de productie van... more Het doel van dit project was de realisatie van een proof-of-principle keten voor de productie van astaxanthine als oleoresin uit de alg Haematococcus pluvialis in fotobioreactoren in Nederlandse kassen. Astaxanthine is een sterk antioxidant dat bij kan dragen aan een gezonde voeding voor consumenten. Astaxanthine kan in Nederlandse kassen duurzaam worden geproduceerd (kassen als zonnecollector, gebruik afval CO 2 uit industrie, reinigen afvalwater). Binnen dit project is onderzoek verricht naar de selectie van natuurlijke algenstammen op kasniveau, klassieke stamverbetering op labniveau (uitgangsmateriaal), procesverbetering in alle fases van de productie (voorkweek, opkweek, groene fase, rode fase) door optimalisatie teeltfactoren en hygiënisatie (teelt en productie) en het maken van een proof-of-principle eindproduct (eindformulering) volgens marktspecificatie (markt en economie). Met het project is kennis vergaard om een nieuw verdienmodel voor de productie van een hoogwaardige stof (astaxanthine) in de tuinbouw te realiseren. Er is een proof-of-principle van een economisch rendabele productieketen van uitgangsmateriaal over productie tot product eindformulering volgens specificaties van de markt aangetoond voor de productie van astaxanthine uit Haematococcus pluvialis.