Production of Recombinant -Hexosaminidase A, a Potential Enzyme for Replacement Therapy for Tay-Sachs and Sandhoff Diseases, in the Methylotrophic Yeast Ogataea minuta Hiromi Akeboshi, Yasunori Chiba,* Yoshiko Kasahara, Minako Takashiba, Yuki Takaoka, Mai Ohsawa, Youichi Tajima, Ikuo Kawashima, Daisuke Tsuji, Kohji Itoh, Hitoshi Sakuraba, and Yoshifumi Jigami* Research Center for Glycoscience, ...

A facultatively methylotrophic Mycobacterium was isolated from Cleveland Harbor, Ohio, USA. The isolate, designated ID-Y, used a wide range of carbon and energy sources including methane and several other hydrocarbons. It displayed a growth cycle from rod-shaped exponential-phase cells, with many cell pairs exhibiting V-formation, to cocco-bacillary stationary-phase cells. A fixation technique ...

The Marburg strain of Butyribacterium methylotrophicum did not grow on CO alone but did consume CO during growth on a variety of substrates in the presence of a 100% CO gas phase. We selected a strain (the CO strain) that grew vigorously on CO alone. The ability of the CO strain to grow on CO was stable through multiple transfers in the absence of CO. CO dehydrogenase activity was lower in the ...

The dramatic expansion of the peroxisomal compartment known to occur in the methanol-utilizing yeast Hansenula polymorpha on transfer from glucose- to methanol-containing media was shown to be accompanied by the synthesis of at least six major polypeptides that dominate the polypeptide pattern of total cell extracts analyzed by NaDodSO(4)/polyacrylamide gel electrophoresis. Two of these polypep...

Root nodule isolates from a shrubby legume, Lotononis bainesii, were characterized by 16S rRNA gene sequencing and morphologically by substrate utilization patterns. The symbiotic genome of these isolates was analysed by partial sequencing of the nifH gene. Based on the results of numerical taxonomy, the isolates formed a closely related cluster, showing no correspondence to any of the known rh...

The possibility of using femtosecond pulses from an X-ray free-electron laser to collect diffraction data from protein crystals formed in their native cellular organelle has been explored. X-ray diffraction of submicrometre-sized alcohol oxidase crystals formed in peroxisomes within cells of genetically modified variants of the methylotrophic yeast Hansenula polymorpha is reported and character...

Nitrate analysis in water is one of the most frequently applied methods in environmental chemistry. Current methods for nitrate are generally based on toxic substances. Here, we show that a viable alternative method is to use the enzyme nitrate reductase. The key to applying this Green Chemistry solution for nitrate analysis is plentiful, inexpensive, analytical grade enzyme. We demonstrate tha...

A single-stranded DNA (ssDNA)-binding protein (SSB) that binds to specific upstream sequences of alcohol oxidase (AOX1) promoter of the methylotrophic yeast Pichia pastoris has been isolated and identified as zeta crystallin (ZTA1). The cDNA encoding P.pastoris ZTA1 (PpZTA1) was cloned into an Escherichia coli expression vector, the recombinant PpZTA1 was expressed and purified from E.coli cell...

Biopharmaceuticals are an integral part of modern medicine and pharmacy. Both, the development and the biotechnological production of biopharmaceuticals are highly cost-intensive and require suitable expression systems. In this review we discuss established and emerging tools for reengineering the methylotrophic yeast Pichia pastoris for biopharmaceutical production. Recent advancements of this...

Strain F31T was isolated from meadow grass (Poa trivialis L.) sampled from the city park in Helsinki. Analysis of phenotypic and genotypic properties showed the strain to be related to the group of obligately methylotrophic non-methane utilizing bacteria (methylobacteria) with the ribulose monophosphate pathway of formaldehyde assimilation. Phylogenetic analysis showed the strain to be closely ...