• 한국미생물·생명공학회지
• /
• 제50권1호
• /
• pp.81-88
• /
• 2022

Acetic acid bacteria (AAB) are versatile organisms involved in the production of variety of fermented foods, such as vinegar and kombucha, and products of biotechnological relevance, such as bacterial cellulose. In the present study, Malatya apricot, a variety with protected designation of origin (PDO), and vinegar samples produced using various fruits were used to isolate AAB. The 19 AAB isolates obtained were typed using (GTG)5 fingerprinting, and the ones selected were identified by sequencing either 16S rDNA alone or in combination with 16S-23S rRNA internal transcribed spacer region or ligA gene. While all apricot isolates (n = 10) were Gluconobacter cerinus, vinegar isolates (n = 9) were composed of Komagataeibacter saccharivorans, Acetobacter syzygii, and possible two new species of AAB, Komagataeibacter sp., and Gluconobacter sp. (GTG)5 fingerprinting showed the presence of several genotypes of G. cerinus in the apricot samples. Screening for some technologically relevant properties, including thermotolerance, ethanol tolerance, and cellulose production capability, showed that all Komagataeibacter and some Gluconobacter isolates could tolerate the temperature of 35℃, and that vinegar isolates could tolerate up to 8% ethanol. One isolate, Komagataeibacter sp. GUS3 produced bacterial cellulose (1 g/l) and has the potential to be used for cellulose production.

• Journal of Microbiology and Biotechnology
• /
• 제29권4호
• /
• pp.617-624
• /
• 2019

Bacterial nanocellulose (BNC) which is generally synthesized by several species of bacteria has a wide variety of industrial uses, particularly in the food and material industries. However, the low levels of BNC production during the fermentation process should be overcome to reduce its production cost. Therefore, in this study, we screened and identified a new cellulose-producing bacterium, optimized production of the cellulose, and investigated the morphological properties of the cellulosic materials. Out of 147 bacterial isolates from ripened fruits and traditional vinegars, strain SFCB22-18 showed the highest capacity for BNC production and was identified as Komagataeibacter sp. based on 16S rRNA sequence analysis. During 6-week fermentation of the strain using an optimized medium containing 3.0% glucose, 2.5% yeast extract, 0.24% acetic acid, 0.27% $Na_2HPO_4$, and 0.5% ethanol at $30^{\circ}C$, about 5 g/l of cellulosic material was produced. Both imaging and IR analysis proved that the produced cellulose would be nanoscale bacterial cellulose.

• Journal of Microbiology and Biotechnology
• /
• 제31권3호
• /
• pp.429-438
• /
• 2021

Bacterial cellulose (BC) is widely used in the food industry for products such as nata de coco. The mechanical properties of BC hydrogels, including stiffness and viscoelasticity, are determined by the hydrated fibril network. Generally, Komagataeibacter bacteria produce gluconic acids in a glucose medium, which may affect the pH, structure and mechanical properties of BC. In this work, the effect of pH buffer on the yields of Komagataeibacter hansenii strain ATCC 53582 was studied. The bacterium in a phosphate and phthalate buffer with low ionic strength produced a good BC yield (5.16 and 4.63 g/l respectively), but there was a substantial reduction in pH due to the accumulation of gluconic acid. However, the addition of gluconic acid enhanced the polymer density and mechanical properties of BC hydrogels. The effect was similar to that of the bacteria using glycerol in another carbon metabolism circuit, which provided good pH stability and a higher conversion rate of carbon. This study may broaden the understanding of how carbon sources affect BC biosynthesis.

• Journal of Microbiology and Biotechnology
• /
• 제30권9호
• /
• pp.1430-1435
• /
• 2020

Bacterial cellulose (BC) has outstanding physical and chemical properties, including high crystallinity, moisture retention, and tensile strength. Currently, the major producer of BC is Komagataeibacter xylinus. However, due to limited tools of expression, this host is difficult to engineer metabolically to improve BC productivity. In this study, a regulated expression system for K. xylinus with synthetic ribosome binding site (RBS) was developed and used to engineer a BC biosynthesis pathway. A synthetic RBS library was constructed using green fluorescent protein (GFP) as a reporter, and three synthetic RBSs (R4, R15, and R6) with different strengths were successfully isolated by fluorescence-activated cell sorting (FACS). Using synthetic RBS, we optimized the expression of three homologous genes responsible for BC production, pgm, galU, and ndp, and thereby greatly increased it under both static and shaking culture conditions. The final titer of BC under static and shaking conditions was 5.28 and 3.67 g/l, respectively. Our findings demonstrate that reinforced metabolic flux towards BC through quantitative gene expression represents a practical strategy for the improvement of BC productivity.

• Journal of Microbiology and Biotechnology
• /
• 제32권8호
• /
• pp.967-975
• /
• 2022

Kombucha mutualistic community (KMC) is composed by acetic acid bacteria and yeasts, producing fermented tea with health benefits. As part of the BIOlogy and Mars EXperiment (BIOMEX) project, the effect of Mars-like conditions on the KMC was analyzed. Here, we analyzed metagenome-assembled genomes (MAGs) of the Komagataeibacter, which is a predominant genus in KMC, to understand their roles in the KMC after exposure to Mars-like conditions (outside the International Space Station) based on functional genetic elements. We constructed three MAGs: K. hansenii, K. rhaeticus, and K. oboediens. Our results showed that (i) K. oboediens MAG functionally more complex than K. hansenii, (ii) K. hansenii is a keystone in KMCs with specific functional features to tolerate extreme stress, and (iii) genes related to the PPDK, betaine biosynthesis, polyamines biosynthesis, sulfate-sulfur assimilation pathway as well as type II toxin-antitoxin (TA) system, quorum sensing (QS) system, and cellulose production could play important roles in the resilience of KMC after exposure to Mars-like stress. Our findings show the potential mechanisms through which Komagataeibacter tolerates the extraterrestrial stress and will help to understand minimal microbial composition of KMC for space travelers.

• Journal of Microbiology and Biotechnology
• /
• 제31권10호
• /
• pp.1366-1372
• /
• 2021

Bacterial nanocellulose (BNC) is a biocompatible material with a lot of potential. To make BNC commercially feasible, improvements in its production and surface qualities must be made. Here, we investigated the in situ fermentation and generation of BNC by addition of different cellulosic substrates such as Avicel and carboxymethylcellulose (CMC) and using Komagataeibacter sp. SFCB22-18. The addition of cellulosic substrates improved BNC production by a maximum of about 5 times and slightly modified its structural properties. The morphological and structural properties of BNC were investigated by using Fourier transform-infrared spectroscopy (FT-IR), scanning electron microscopy and X-ray diffraction. Furthermore, a type-A cellulose-binding protein derived from Clostridium thermocellum, CtCBD3, was used in a novel biological analytic approach to measure the surface crystallinity of the BNC. Because Avicel and CMC may adhere to microfibrils during BNC synthesis or crystallization, cellulose-binding protein could be a useful tool for identifying the crystalline properties of BNC with high sensitivity.

• Journal of Species Research
• /
• 제12권spc2호
• /
• pp.7-14
• /
• 2023

Several genera belonging to the family Acetobacteraceae were generally considered to be acetic acid bacteria (AAB) which produce acetic acid from alcohols using an oxidation pathway. Some species of the family Acetobacteraceae have been of interest to the industry due to their capability to produce vinegar. In 2018-2020, several bacterial strains were isolated from plants, fruits and vinegar in various regions of the Republic of Korea. Based on the 16S rRNA gene sequences, seven species classified into the family Acetobacteraceae were found to be ones unrecorded in the Republic of Korea, including two Asaia species, one Gluconacetobacter species, three Gluconobacter species and one Komagataeibacter species. As a result, we report Asaia lannensis AF11C3 (=KACC 22050) isolated from plant Chrysanthemum zawadskii, Asaia platycodi AF15C2 (=KACC 22051) isolated from plant Isodon inflexus, Gluconacetobacter liquefaciens C23-3 (=KACC 22064) isolated from fruit of Pyrus pyrifolia, Gluconobacter cerinus BGF2-R2 (=KACC 22053) isolated from fruit of Prunus armeniaca, Gluconobacter kondonii FR39A4 (=KACC 22388) isolated from persimmon fruit, Gluconobacter thailandicus FR36C4 (=KACC 22057) isolated from fruit of Pseudocydonia sinensis and Komagataeibacter melaceti SPV-2 (=KACC 22058) isolated from potato vinegar.

• Toxicological Research
• /
• 제35권1호
• /
• pp.45-63
• /
• 2019

In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ($BMM{\Phi}$) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.