Dataset Information

Production of bacterial cellulose by Gluconacetobacter hansenii UAC09 using coffee cherry husk.

ABSTRACT: The work is aimed to investigate the suitability of underutilized coffee cherry husk (CCH) for the production and optimization of bacterial cellulose (BC) by Gluconacetobacter hansenii UAC09 and to study the physico-mechanical properties of BC films. CCH extract was used as a carbon source in various concentrations along with other nutritional components such as nitrogen (corn steep liquor, urea) and additives (ethyl alcohol, acetic acid). Concentration of CCH extract at 1:1 (w/v) along with 8% (v/v) corn steep liquor, 0.2% (w/v) urea, combination of 1.5% ethyl alcohol and 1.0% (v/v) acetic acid resulted in the production of 5.6-8.2 g/L of BC. BC had tensile strength varying between 28.5 and 42.4 MPa. BC produced with CCH and Hestrin and Schramm (HS) media did not differ in structure as analyzed by FT-IR. Scanning electron microscopic studies indicated BC to contain reticulated network of fine fibers. Under optimized condition, based on the other additives, CCH produced more than three folds yield of BC (5.6-8.2 g/L) than control medium (1.5 g/L). This is the first report on the use of CCH for the production of BC and paved way for the utilization of organic wastes with pectin and high polyphenol content.

SUBMITTER: Rani MU

PROVIDER: S-EPMC3671040 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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Production of bacterial cellulose by Gluconacetobacter hansenii UAC09 using coffee cherry husk.

Rani M Usha MU Appaiah K A Anu KA

Journal of food science and technology 20110528 4

The work is aimed to investigate the suitability of underutilized coffee cherry husk (CCH) for the production and optimization of bacterial cellulose (BC) by Gluconacetobacter hansenii UAC09 and to study the physico-mechanical properties of BC films. CCH extract was used as a carbon source in various concentrations along with other nutritional components such as nitrogen (corn steep liquor, urea) and additives (ethyl alcohol, acetic acid). Concentration of CCH extract at 1:1 (w/v) along with 8% ...[more]

PMID: 24425978

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Project description:Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 Å for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. The results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the hypothesis that it is the extrusion mechanism and order in linearly arrayed TCs that enables production of crystalline cellulose.

Project description: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 that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of peptidoglycan in the assembly of crystalline cellulose.

Dataset Information

Production of bacterial cellulose by Gluconacetobacter hansenii UAC09 using coffee cherry husk.

Publications

Production of bacterial cellulose by Gluconacetobacter hansenii UAC09 using coffee cherry husk.

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