Cloning and characterization of a pyridoxine 5′‐phosphate oxidase from silkworm,<i> Bombyx mori</i>

Huang, ShuoHao; Shi, Renyi; Zhang, J. Y.; Wang, Z.; Huang, LongQuan

doi:10.1111/j.1365-2583.2009.00880.x

Cited by 10 publications

(2 citation statements)

References 19 publications

(23 reference statements)

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“…Since B. mori is a large silk-secreting insect, its immense protein turnover needs the timely support of PLP. In order to understand the metabolic mechanism of VB6 in B. mori, the cDNAs encoding PLK and PNPO in B. mori larvae were cloned (Shi et al, 2007;Huang et al, 2009). In this study, the recombinant B. mori PLK was expressed in E. coli as a fusion protein with a hexa-histidine affinity tag, purified by Ni Sepharose affinity column chromatography and characterized.…”

Section: Introductionmentioning

confidence: 99%

Recombinant expression, purification and characterization of Bombyx mori (Lepidoptera: Bombycidae) pyridoxal kinase

Huang¹,

Ma²,

Zhang³

et al. 2011

Eur. J. Entomol.

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Abstract. Pyridoxal kinase (PLK; EC 2.7.1.35) is a key enzyme in the metabolism of vitamin B6 (VB6) in Bombyx mori. A fusion expressional vector pET-22b-BPLK-His was constructed using a sub-cloning technique, the recombinant B. mori PLK was then expressed in Escherichia coli, purified and characterized. Bioinformatics were used to deduce the protein structure and genomic organization of this enzyme. Using Ni Sepharose affinity column chromatography, the recombinant protein was purified to very high degree (approximately 90%). The recombinant PLK exhibits a high specific enzymatic activity (1800 nmol/min/mg of protein). The maximum catalytic activity of this enzyme was recorded over a narrow pH range (5.5-6.0) and Zn 2+ is the most effective cation for catalysis under saturating substrate concentrations. When only triethanolamine is present as the cation, K + is an activator of PLK. A double reciprocal plot of initial velocity suggests that the enzyme catalyses the reaction by means of a sequential catalytic mechanism. Under optimal conditions, the Km value for the substrates of ATP and pyridoxal are 57.9 ± 5.1 and 44.1 ± 3.9 µM. B. mori's genome contains a single copy of the PLK gene, which is 7.73 kb long and contains five exons and four introns, and is located on the eighth chromosome. The PLK may be a dimer with two identical subunits under native conditions, and it is hypothesized that each monomer contains eight -helices ( 1-8), nine -strands ( 1-9) and two segments of 310 helices.

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Section: Introductionmentioning

confidence: 99%

Recombinant expression, purification and characterization of Bombyx mori (Lepidoptera: Bombycidae) pyridoxal kinase

Huang¹,

Ma²,

Zhang³

et al. 2011

Eur. J. Entomol.

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“…Though experimental evidence is meager, PNPO is involved in phenazine biosynthesis (Domitrovic et al, 2015; Musayev et al, 2003). It binds with flavin mononucleotide and oxidizes pyridoxine 5′‐phosphate or pyridoxamine 5′‐phosphate to form pyridoxal 5′‐phosphate, playing a pivotal role in cell metabolism (Blankenfeldt & Parsons, 2014; Diederich et al, 2017; Huang et al, 2009). PNPO catalyzes oxidation‐reduction reactions that produce hydrogen peroxide along with pyridoxal 5′‐phosphate (Sang et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Dipteran endoparasitoid Exorista bombycis utilizes antihemocyte components against host defense of silkworm Bombyx mori

Moteriya

Hungund²,

Pradeep

2022

Arch Insect Biochem Physiol

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Dipteran endoparasitoids avoid host immune response; however, antidefense components from the Dipterans are unknown. Infestation of commercial silkworm Bombyx mori Linnaeus (Lepidoptera: Bombycidae) by endoparasitoid Exorista bombycis Louis (Diptera: Tachinidae) induced immune reactions, cytotoxicity, granulation, degranulation, and augmented release of cytotoxic marker enzyme lactate dehydrogenase (LDH), and degranulation‐mediator enzyme β‐hexosaminidase in hemocytes. In this study, by reverse phase high‐performance liquid chromatography, fractions of E. bombycis larval tissue protein with antihemocytic activity are separated. From the fraction, peptides of hemocyte aggregation inhibitor protein (HAIP) and pyridoxamine phosphate oxidase (PNPO) are identified by mass spectrometry. Interacting partners of HAIP and PNPO are retrieved that further enhance the virulence of the parasitoid. PNPO and HAIP genes showed a four‐ to seven fold increase in expression in the integument of the parasitoid larva. Together, the dipteran endoparasitoid E. bombycis exploit antihemocyte activity to inhibit host defense reactions in addition to defense evasion contemplated.

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Vitamin B6 salvage enzymes: Mechanism, structure and regulation

Salvo

Contestabile

Safo

2011

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

208

188

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Cloning and characterization of a pyridoxine 5′‐phosphate oxidase from silkworm, Bombyx mori

Cited by 10 publications

References 19 publications

Recombinant expression, purification and characterization of Bombyx mori (Lepidoptera: Bombycidae) pyridoxal kinase

Recombinant expression, purification and characterization of Bombyx mori (Lepidoptera: Bombycidae) pyridoxal kinase

Dipteran endoparasitoid Exorista bombycis utilizes antihemocyte components against host defense of silkworm Bombyx mori

Vitamin B6 salvage enzymes: Mechanism, structure and regulation

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