Bacterial cellulose production by Komagataeibacter hansenii can be improved by successive batch culture.

Bacterial cellulose (BC) is a biopolymer principally synthetized by strains of the genus Komagataeibacter. However, high costs and low production yield make large-scale application difficult. The aim of this work was to evaluate the effects of successive batch culture before fermentation on the ability to increase the capacity of bacterial cellulose biosynthesis by a low-producing strain. The Komagataeibacter hansenii strain ATCC 23,769 was initially cultivated in fermentation broth for two periods of 35 or 56 days under static conditions. At the end of each period of time, they were transferred to new broth to be cultivated again (new batch culture cycle) for 35 or 56 days and carried out in parallel with a 10-day fermentation to determine the quantity of BC produced. As a result, a greater increase was observed after the end of the second and third batch cultures of 56 days (increases of 137% and 187% in relation to the nonbatch cultured strain, respectively). The produced samples presented higher crystallinity and thermal properties but lower water holding capacity. Through this work, it was concluded that the longer the batch culture time was, the greater the increase in the capacity of cellulose biosynthesis, which also depended on the number of successive batch culture cycles carried out.

Nutritional Supplementation with Amino Acids on Bacterial Cellulose Production by Komagataeibacter intermedius: Effect Analysis and Application of Response Surface Methodology.

Bacterial cellulose (BC) is a biopolymer mainly produced by acetic acid bacteria (AAB) that has several applications in the medical, pharmaceutical, and food industries. As other living organisms, AAB require sources of chemical elements and nutrients, which are essential for their multiplication and metabolite production. So, the knowledge of the nutritional needs of microorganisms that have important industrial applications is necessary for the nutrients to be supplied in the appropriate form and amount. Considering that the choice of different nutrients as nitrogen source can result in different metabolic effects, this work aimed to verify the effects of amino acid supplementation in the culture media for BC production by an AAB strain (Komagataeibacter intermedius V-05). For this, nineteen amino acids were tested, selected, and optimized through a Plackett and Burman factorial design and central composite design to determine the optimal concentrations of each required amino acid. Membranes produced under optimal conditions were characterized in relation to chemical structure and properties by X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), infrared spectroscopy (FT-IR), and hydrophilic properties. Three amino acids had a significant positive effect and were required: aspartic acid (1.5 g L-1), phenylalanine (1.5 g L-1), and serine (3.0 g L-1). Conversely, all sulfur and positively charged amino acids had a negative effect and reduced the production yield. After optimization and validation steps, a production level of 3.02 g L-1 was achieved. Membranes produced from optimized media by this strain presented lower crystallinity index but greater thermal and hydrophilic properties than those produced from standard HS medium.

Komagataeibacter intermedius V-05: An Acetic Acid Bacterium Isolated from Vinegar Industry, with High Capacity for Bacterial Cellulose Production in Soybean Molasses Medium.

RESEARCH BACKGROUND: Despite the great properties of bacterial cellulose, its manufacture is still limited due to difficulties in large-scale production. These problems are mainly related to low production yields and high overall costs of the conventional culture media normally used. To surpass these problems, it is necessary to identify new cheap and sustainable carbon sources. Thus, this work aims to isolate and select a high cellulose-producing Komagataeibacter strain from vinegar industry, and study its potential for bacterial cellulose synthesis in an industrial soybean co-product, known as soybean molasses, used as fermentation medium. EXPERIMENTAL APPROACH: One isolated strain was able to produce high amount of cellulose in the standard Hestrin-Schramm medium, so we tested its ability to produce this biopolymer in a soybean molasses medium. The characteristics and properties of the produced bacterial cellulose membranes were analyzed by thermogravimetric analysis, X-ray diffraction, infrared spectroscopy, water-holding capacity and rehydration ratio. Genetic analysis of the selected strain served to determine its genus and species. RESULTS AND CONCLUSIONS: An isolated strain that produced the highest amount of cellulose in Hestrin-Schramm medium (3.7 g/L) was genetically identified as Komagataeibacter intermedius V-05. This strain produced 10.0 g/L of cellulose in soybean molasses medium. Membranes from both substrates had similar chemical structure, crystallinity and thermal degradation. Soybean molasses proved to be a suitable alternative medium for biosynthesis of cellulose in comparison with the standard medium. In addition to providing higher production yield, the membranes showed great structural characteristics, similar to those obtained from standard medium. NOVELTY AND SCIENTIFIC CONTRIBUTION: In this research, we have isolated and identified a Komagataeibacter strain which exhibits a high capacity for cellulose production in soybean molasses. The isolation and selection of strains with high capacity for microbial metabolite production is important for decreasing bioprocess costs. Furthermore, as there is a necessity today to find cheaper carbon sources to obtain microbial products at a lower cost, soybean molasses represents an interesting alternative medium to produce bacterial cellulose for its industrial application.

Acetic Acid Fermentation of Soybean Molasses and Characterisation of the Produced Vinegar.

Soybean molasses is a by-product from the production of protein concentrate from soybean meal that predominantly contains sugars, with sucrose as the major component. In Brazil, soybean molasses is used for animal feed or it is discarded, although some industries use it to produce ethanol. This study aims to evaluate the parameters required for the acetic acid fermentation of soybean molasses, and characterise the resultant vinegar. To study the most suitable parameters for the acetic acid fermentation, vinegar was produced from the alcohol fermentation of soybean molasses through eight fermentation cycles: five for adaptation and three for production. The average acidity of the acetic acid fermentation product was 50.60 g/L, with an acetic acid fermentation yield, total yield of acetic acid in broth and productivity 65.01%, 92.76% and 0.033 g/(L·h), respectively. The vinegar produced from soybean molasses has an acidity of 5.07% (m/V), residual ethanol content 0.17% (m/V), sugars 7.86% (m/V), dry extract 14.67% (m/V), ash 2.27% (m/V) and a density of 1.023 g/cm3. The contents of total phenolics and isoflavones decreased after the alcohol and acetic acid fermentations. Moreover, the isoflavones profile of the fermented product comprised only three forms: daidzein, glycitin and genistin. According to our results, 3460 L of vinegar can be produced for every tonne of soy molasses, with an acetic acid concentration of 40 g/L, the minimum required by the legislation on vinegar production. Thus, these findings demonstrate that soy molasses represents a useful raw material for the production of vinegar.

Acetic Acid Bacteria in the Food Industry: Systematics, Characteristics and Applications.

The group of Gram-negative bacteria capable of oxidising ethanol to acetic acid is called acetic acid bacteria (AAB). They are widespread in nature and play an important role in the production of food and beverages, such as vinegar and kombucha. The ability to oxidise ethanol to acetic acid also allows the unwanted growth of AAB in other fermented beverages, such as wine, cider, beer and functional and soft beverages, causing an undesirable sour taste. These bacteria are also used in the production of other metabolic products, for example, gluconic acid, l-sorbose and bacterial cellulose, with potential applications in the food and biomedical industries. The classification of AAB into distinct genera has undergone several modifications over the last years, based on morphological, physiological and genetic characteristics. Therefore, this review focuses on the history of taxonomy, biochemical aspects and methods of isolation, identification and quantification of AAB, mainly related to those with important biotechnological applications.