BB-kb@ldeleger:BB-kb-11437594 / 86-1277 JSONTXT

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Id Subject Object Predicate Lexical cue NCBI_Taxonomy OntoBiotope
T2 0-1191 Paragraph denotes Hexose oxidase (D-hexose:O(2)-oxidoreductase, EC 1.1.3.5, HOX) normally found in the red alga Chondrus crispus was produced heterologously in different host systems. Full-length HOX polypeptide was produced in Escherichia coli, but no HOX activity could be detected. In contrast, active HOX could be produced in the methylotrophic yeast Pichia pastoris. Several growth physiological and genetic approaches for optimization of hexose oxidase production in P. pastoris were investigated. Our results indicate that specific growth conditions are essential in order to produce active HOX with the correct conformation. Furthermore, HOX seems to be activated by proteolytic cleavage of the full-length polypeptide chain into two fragments, which remain physically associated. Attempts to direct HOX to the extracellular compartment using the widely used secretion signals from Saccharomyces cerevisiae invertase or alpha-mating factor failed. However, we show in this study that HOX is transported out of P. pastoris via a hitherto unknown mechanism and that it is possible to enhance this secretion by mutagenesis from below the detection limit to at least 250 mg extracellular enzyme per liter.
T5 85-110 Habitat denotes red alga Chondrus crispus OBT:002758
T6 210-226 Microorganism denotes Escherichia coli 562
T7 235-247 Phenotype denotes HOX activity OBT:000371
T8 316-330 Phenotype denotes methylotrophic OBT:000734
T9 337-352 Microorganism denotes Pichia pastoris 4922
E1 337-352 Exhibits denotes Pichia pastoris
T10 426-451 Phenotype denotes hexose oxidase production OBT:000371
T11 455-466 Microorganism denotes P. pastoris 4922
T12 872-896 Microorganism denotes Saccharomyces cerevisiae 4932
T13 910-922 Phenotype denotes alpha-mating OBT:000173
T14 1000-1011 Microorganism denotes P. pastoris 4922
R1 E1 T9 Microorganism Pichia pastoris,Pichia pastoris
R2 E1 T8 Property Pichia pastoris,methylotrophic