Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity

PloS One
Ying DengTeh-hui Kao

Abstract

Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC). These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed...Continue Reading

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Citations

Mar 19, 2019·Microbial Biotechnology·Paulina JacekStanisław Bielecki
Apr 16, 2021·Applied Microbiology and Biotechnology·Philipp Moritz FrickeTino Polen
Oct 12, 2021·FEMS Microbiology Reviews·Wiem AbidiPetya Violinova Krasteva

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Datasets Mentioned

BETA
EFG85846
EFG85940

Methods Mentioned

BETA
X-ray
NMR
scanning electron microscopy
Polymerase Chain Reactions
PCR
PCRs

Software Mentioned

BLASTN
TOSS
BLASTtx
CP

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