BB-norm+ner@ldeleger:BB-norm+ner-11437594 JSONTXT

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Id Subject Object Predicate Lexical cue NCBI_Taxonomy OntoBiotope
T1 0-85 Title denotes Optimization of the production of Chondrus crispus hexose oxidase in Pichia pastoris.
T3 34-50 Habitat denotes Chondrus crispus OBT:002758
T4 69-84 Microorganism denotes Pichia pastoris 4922
T2 86-1277 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 171-196 Habitat denotes red alga Chondrus crispus OBT:002758
T6 296-312 Microorganism denotes Escherichia coli 562
T7 321-333 Phenotype denotes HOX activity OBT:000371
T8 402-416 Phenotype denotes methylotrophic OBT:000734
T9 423-438 Microorganism denotes Pichia pastoris 4922
T10 512-537 Phenotype denotes hexose oxidase production OBT:000371
T11 541-552 Microorganism denotes P. pastoris 4922
T12 958-982 Microorganism denotes Saccharomyces cerevisiae 4932
T13 996-1008 Phenotype denotes alpha-mating OBT:000173
T14 1086-1097 Microorganism denotes P. pastoris 4922