This
is the first study on improving lactobionic acid
(LBA) production capacity in Pseudomonas taetrolens by genetic engineering. First, quinoprotein glucose dehydrogenase
(GDH) was identified as the lactose-oxidizing enzyme of P. taetrolens. Of the two types of GDH genes in P. taetrolens, membrane-bound (GDH1) and soluble
(GDH2), only GDH1 showed lactose-oxidizing activity. Next, the genetic
tool system for P. taetrolens was developed
based on the pDSK519 plasmid for the first time, and GDH1 gene was
homologously expressed in P. taetrolens. Recombinant expression of the GDH1 gene enhanced intracellular
lactose-oxidizing activity and LBA production of P.
taetrolens in flask culture. In batch fermentation
of the recombinant P. taetrolens using
a 5 L bioreactor, the LBA productivity of the recombinant P. taetrolens was approximately 17% higher (8.70
g/(L h)) than that of the wild type (7.41 g/(L h)). The LBA productivity
in this study is the highest ever reported using bacteria as production
strains for LBA.
Coptis chinensis (CC) is used in conventional Chinese medicine. The main active components of CC are isoquinoline alkaloids, including berberine, coptisine, palmatine, and magnoflorine; all these are known to have several pharmacological properties. Poly(vinyl alcohol) (PVA) is a well-known synthetic biocompatible polymer suitable for a range of pharmaceutical uses; it can be used as a matrix for the incorporation of functional materials and has a wide range of applications in the cosmetics, food, pharmaceutical, and packaging industries. In this study, PVA-based electrospun nanofibers containing CC extract were successfully fabricated. Furthermore, the effects of different CC extract contents on the morphologies, and antimicrobial and antifungal properties of PVA/CC extract nanofibers were investigated. Morphological changes were observed using different molecular weights of PVA. For characterization, field-emission scanning electron microscopy, thermogravimetric analysis, and Fourier transform infrared analysis were performed. The effectiveness of these nanofibers has been demonstrated by evaluating the thermal stability against Staphylococcus aureus, antimicrobial activity against Staphylococcus aureus and Staphylococcus epidermidis, and the antifungal activity against the fungi Aureobasidium pullulans and Penicillium pinophilum. The PVA/CC extract nanofibers were found to have excellent antibacterial and antifungal activity and thermal stability; hence, their use in medicinal sectors is highly recommended.
Highlights
Glucose dehydrogenase from
Pseudomonas taetrolens
could produce maltobionic acid.
The glucose dehydrogenase gene was homologously expressed in
P. taetrolens
.
Maltose-oxidizing activity and MBA production titer of
P. taetrolens
was improved.
MBA productivity of the recombinant
P. taetrolens
was increased to 9.52 g/L/h.
High-level production of MBA from HMCS was achieved by the recombinant
P. taetrolens
.
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