Enhancing small-scale acetification processes using adsorbed Acetobacter pasteurianus UMCC 2951 on κ-carrageenan-coated luffa sponge.

Background: This study explored the utilization of luffa sponge (LS) in enhancing acetification processes. LS is known for having high porosity and specific surface area, and can provide a novel means of supporting the growth of acetic acid bacteria (AAB) to improve biomass yield and acetification rate, and thereby promote more efficient and sustainable vinegar production. Moreover, the promising potential of LS and luffa sponge coated with κ-carrageenan (LSK) means they may represent effective alternatives for the co-production of industrially valuable bioproducts, for example bacterial cellulose (BC) and acetic acid. Methods: LS and LSK were employed as adsorbents for Acetobacter pasteurianus UMCC 2951 in a submerged semi-continuous acetification process. Experiments were conducted under reciprocal shaking at 1 Hz and a temperature of 32 °C. The performance of the two systems (LS-AAB and LSK-AAB respectively) was evaluated based on cell dry weight (CDW), acetification rate, and BC biofilm formation. Results: The use of LS significantly increased the biomass yield during acetification, achieving a CDW of 3.34 mg/L versus the 0.91 mg/L obtained with planktonic cells. Coating LS with κ-carrageenan further enhanced yield, with a CDW of 4.45 mg/L. Acetification rates were also higher in the LSK-AAB system, reaching 3.33 ± 0.05 g/L d as opposed to 2.45 ± 0.05 g/L d for LS-AAB and 1.13 ± 0.05 g/L d for planktonic cells. Additionally, BC biofilm formation during the second operational cycle was more pronounced in the LSK-AAB system (37.0 ± 3.0 mg/L, as opposed to 25.0 ± 2.0 mg/L in LS-AAB). Conclusions: This study demonstrates that LS significantly improves the efficiency of the acetification process, particularly when enhanced with κ-carrageenan. The increased biomass yield, accelerated acetification, and enhanced BC biofilm formation highlight the potential of the LS-AAB system, and especially the LSK-AAB variant, in sustainable and effective vinegar production. These systems offer a promising approach for small-scale, semi-continuous acetification processes that aligns with eco-friendly practices and caters to specialized market needs. Finally, this innovative method facilitates the dual production of acetic acid and bacterial cellulose, with potential applications in biotechnological fields.

Modeling Textural Properties of Cooked Germinated Brown Rice Using the near-Infrared Spectra of Whole Grain.

If a non-destructive and rapid technique to determine the textural properties of cooked germinated brown rice (GBR) was developed, it would hold immense potential for the enhancement of the quality control process in large-scale commercial rice production. We combined the Fourier transform near-infrared (NIR) spectral data of uncooked whole grain GBR with partial least squares (PLS) regression and an artificial neural network (ANN) for an evaluation of the textural properties of cooked germinated brown rice (GBR); in addition, data separation and spectral pretreatment methods were investigated. The ANN was outperformed in the evaluation of hardness by a back extrusion test of cooked GBR using the smoothing combined with the standard normal variate pretreated NIR spectra of 188 whole grain samples in the range of 4000-12,500 cm-1. The calibration sample set was separated from the prediction set by the Kennard-Stone method. The best ANN model for hardness, toughness, and adhesiveness provided R2, r2, RMSEC, RMSEP, Bias, and RPD values of 1.00, 0.94, 0.10 N, 0.77 N, 0.02 N, and 4.3; 1.00, 0.92, 1.40 Nmm, 9.98 Nmm, 1.6 Nmm, and 3.5; and 0.97, 0.91, 1.35 Nmm, 2.63 Nmm, -0.08 Nmm, and 3.4, respectively. The PLS regression of the 64-sample KDML GBR group and the 64-sample GBR group of various varieties provided the optimized models for the hardness of the former and the toughness of the latter. The hardness model was developed by using 5446.3-7506 and 4242.9-4605.4 cm-1, which included the amylose vibration band at 6834.0 cm-1, while the toughness model was from 6094.3 to 9403.8 cm-1 and included the 6834.0 and 8316.0 cm-1 vibration bands of amylose, which influenced the texture of the cooked rice. The PLS regression models for hardness and toughness had the r2 values of 0.85 and 0.82 and the RPDs of 2.9 and 2.4, respectively. The ANN model for the hardness, toughness, and adhesiveness of cooked GBR could be implemented for practical use in GBR production factories for product formulation and quality assurance and for further updating using more samples and several brands to obtain the robust models.

Evaluation of Precision and Sensitivity of Back Extrusion Test for Measuring Textural Qualities of Cooked Germinated Brown Rice in Production Process.

The textural qualities of cooked rice may be understood as a dominant property and indicator of eating quality. In this study, we evaluated the precision and sensitivity of a back extrusion (BE) test for the texture of cooked germinated brown rice (GBR) in a production process. BE testing of the textural properties of cooked GBR rice showed a high precision of measurement in hardness, toughness and stickiness tests which indicated by the repeatability and reproductivity test but the sensitivity indicated by coefficient of variation of the texture properties. The findings of our study of the effects on cooked GBR texture of different soaking and incubation durations in the production of Khao Dawk Mali 105 (KDML 105) GBR, as measured by BE testing, confirmed that our original protocol for evaluation of the precision and sensitivity of this texture measurement method. The coefficients of determination (R2) of hardness, toughness and stickiness tests and the incubation time at after 48 hours of soaking were 0.82, 0.81 and 0.64, respectively. The repeatability and reproducibility of reliable measurements, which have a low standard deviation of the greatest difference between replicates, are considered to indicate high precision. A high coefficient of variation where relatively wide variations in the absolute value of the property can be detected indicates high sensitivity when small resolutions can be detected, and vice versa. The sensitivity of the BE tests for stickiness, toughness and hardness all ranked higher, in that order, than the sensitivity of the method for adhesiveness, which ranked lowest. The coefficients of variation of these texture parameters were 31.26, 20.59, 19.41 and 18.72, respectively. However, the correlation coefficients among the texture properties obtained by BE testing were not related to the precision or sensitivity of the test. By obtaining these results, we verified that our original protocol for the determination of the precision and sensitivity of food texture measurements which was successfully used for GBR texture measurement.

Impact of temperature on gluconic acid production during acetification by Acetobacter aceti.

The current approach to gluconic acid production is acetification at 30°C, a temperature that can be difficult to maintain in tropical countries. This study investigated the production of gluconic acid during acetification by Acetobacter aceti WK at high temperatures. An acid-tolerant and thermotolerant species, A. aceti WK, was used for acetification at three different temperatures, namely, 30°C (normal temperature), 37°C, and 40°C (high temperature). Acetification was performed in a 100 L bioreactor with 0.15% CaCl2 for protection of the cells against high temperatures. The production of the organic acids, that is, acetic acid, gluconic acid, 2-keto gluconic acid, glucuronic acid, citric acid, succinic acid, lactic acid, and formic acid, was analyzed. Under acetification in the target total concentration of 80 g/L, the highest acetic acid content (39.3 g/L) was obtained at 37°C with an acetification rate of 0.3013 g/L/h, while the acetic acid content and acetification rate achieved at 30°C were 31 g/L and 0.3089 g/L/h, respectively. Additionally, gluconic acid presented at the highest concentration of 2.17 g/L. The rate of production of gluconic acid was 0.0169 g/L/h at 37°C. This acetification process at 37°C will be valuable as an alternative source for gluconic acid production for commercial applications.

Defining Paenibacillus azoreducens (P8) and Acetobacter pasteurianus (UMCC 2951) strains performances in producing acetic acid.

In this study, spore-forming bacteria isolated from saccharified rice were selected for producing acetic acid. From the screening of 15 strains, P8 strain was chosen as a candidate. The strain was identified as Paenibacillus azoreducens by 16S rRNA analysis (99.85% similarity with P. azoreducens CM1T). Acetic acid is the main component of vinegar but also an industrial commodity produced by chemical synthesis. Sustainable routes for obtaining acetic acid are of great interest for decreasing the environmental impact generated by chemical syntheses. Biological acetic acid production is effective for vinegar production by acetic acid bacteria, but it cannot economically compete with the chemical synthesis for producing it as a pure commodity. Considering the need to improve the yield of pure acetic acid produced by microbial conversions, in this study, P8 strain was chosen for designing processes in different fermentation conditions. Tests were conducted in single and semi-continuous systems, using rice wine as substrate. Acetic acid produced by P8 strain was compared with that of Acetobacter pasteurianus (UMCC 2951), a strain known for producing acetic acid from rice wine. Even though the fermentation performances of P. azoreducens P8 were slightly lower than those of acetic acid bacteria usually used for vinegar production, results highlight its suitability for producing acetic acid. The final acetic acid produced by P. azoreducens P8 was 73 g/L, in a single stage fermentation, without losses. In nine cycles of semi-continuous regime the average of acetification rate was 0.814 (g/L/days). Two main attributes of P. azoreducens P8 are of relevance for producing acetic acid, namely the ability to grow at temperature higher (+ 37°C), than mesophilic acetic acid bacteria, and the absence of cytoplasmic assimilation of acetic acid. These features allow to design multiple strains cultures, in which P. azoreducens can acts as a helper strain. Based on our results, the new isolate P. azoreducens P8 can be propagated in fermenting broths for boosting acetic acid production, under the selected conditions, and used in combination with acetic acid bacteria to produce biological acetic acid, as a non-food grade commodity.

Sulphation and Hydrolysis Improvements of Bioactivities, and Immuno-Modulatory Properties of Edible Amanita hemibapha Subspecies javanica (Corner and Bas) Mucilage Polysaccharide as a Potential in Personalized Functional Foods.

In this study, the mucilage polysaccharide (MP) from Amanita hemibapha subspecies javanica was prepared by hot water extraction and ethanol precipitation and then fractionated using anion-exchange chromatography equipped with a DEAE Sepharose fast flow column. The most immune-enhancing polysaccharide fraction 2 (MPF2) was subjected to a structural modification such as hydrolysis or over-sulphation. The sulphate and molecular weight (Mw) of over-sulphated (OS1-3) and hydrolysed (HS1-3) derivatives of MPF2 differed between 9.85% and 14.2% and 32.8 and 88.1 × 103 g/mol, respectively. Further, the immune-enhancing properties of MPF2 and its derivatives were tested on RAW264.7 and NK cells through various in vitro assays. Interestingly, a low molecular weight of HS1-3 significantly increased the nitric oxide (NO) production (p < 0.05) more than MPF2, indicating that Mw is a major factor in RAW264.7 cell stimulation. In addition, RAW264.7 cells produced various cytokines by up-regulating mRNA expression levels and the activation of nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. On the other hand, OS1-3-treated natural killer (NK) cells induced cytotoxicity in HepG2 cells through the expression of IFN-γ, Grandzyme-B, perforin, NKp30, and FasL. These results demonstrated that sulphate derivatives play an important role in NK cell activation. Further, this study also explores how polysaccharide binds to RAW264.7 and NK cells. MPF2 and HS3 may activate RAW264.7 cells via binding to TLR4 receptors, and OS2 could be activated through the CR3 signalling pathways.

Extraction, Structural Characterisation, and Immunomodulatory Properties of Edible Amanitahemibapha subspecies javanica (Corner and Bas) Mucilage Polysaccharide as a Potential of Functional Food.

This research aimed to extract mucilage polysaccharides (MP) from Amanita hemibapha subspecies javanica (Corner and Bas), and further fractionate them using anion-exchange chromatography, yielding two fractions (MPF1 and MPF2). The crude extract, and fractions mainly consisted of carbohydrates (83.5-93.2%) with minor amounts of proteins (5.40-7.20%), and sulphates (1.40-9.30%). Determination of the monosaccharide composition revealed that glucose was the major unit, followed by galactose, mannose, rhamnose, and arabinose. The average molecular weight (MW) of the crude extract and fractions was in the range 104.0-479.4 × 103 g/mol. Interestingly, the crude extract, and fractions did not cause any toxic effect in RAW264.7 cells. However, they stimulated the RAW264.7 cells to release nitric oxide and cytokines through the activation of nuclear factor-kappa B (NF-κB), and mitogen-activated protein kinase (MAPK) pathways via cell surface TLR4. Structural analysis of the most immunestimulating extract fraction, MPF2, revealed that the main backbone consisted of α-D-(1→6)-glucopyranoside. These results suggest that the MPs derived from A. hemibapha subspecies javanica (Corner and Bas) are potent in enhancing immunity; hence, they can be used as a functional ingredient in food products.

Consecutive bacterial cellulose production by luffa sponge enmeshed with cellulose microfibrils of Acetobacter xylinum under continuous aeration.

The bacterial cellulose production (BCP) process, using cellulose microfibrils (CM) of Acetobacter xylinum enmeshed on luffa sponge matrices (LSM) as LSM-CM starter, has been successfully developed where the LSM-CM production process can be recycled through consecutive cycles in limited dissolved oxygen (DO) under continuous aeration. In this study, incremental aeration rates impacted the consecutive cycles. Gluconic acid production, during the process, resulting in the reduction of BCP, was increasingly generated at high aeration from 0.28 to 0.34% at 3 L/min to 0.83-0.97% and 1.52-1.99% at 6 and 9 L/min after 7 d culture at 30 ± 2 °C. To compensate for the negative impact of aeration, 0.10 and 0.15% (w/v) carboxymethyl cellulose (CMC) was found to create a microenvironment for recycled LSM-CM at both high aeration (6 and 9 L/min, respectively). Under nine consecutive BCP cycles, acceptable BC yields (from 5.54 ± 0.5 to 5.89 ± 0.5 g/L) were associated with high biomass at 6 L/min aeration. These results confirm that LSM-CM, combined with CMC called as LSM-CM-CMC, created microenvironments low in DO under high aeration rates and that the recycled LSM-CM-CMC with aeration is an alternative, sustainable, economic process that could be applied for mass BCP.

Increasing the acetification rate of Acetobacter aceti adsorbed on luffa sponge using recycle of incremental oxygenated medium.

Speeding up the production of vinegar from rice wine by acetification, using a packed-bed bioreactor with a luffa sponge matrix (LSM) as adsorption carrier of acetic acid bacteria (AAB), and the effect of oxygenation of the recycled medium were investigated. The 0.06 L/min recycle of medium resulted in a high oxygen-transfer coefficient, while optimal dissolved oxygen (DO) of the medium maximized planktonic AAB cell growth with no contamination due to high acid in an external reservoir without LSM. The highest acetification rate (ETA) of 2.857 ± 0.1 g/L/day was achieved with DO 3.5-4.5 ppm at 35 ± 1 °C. To increase ETA, the optimized oxygenated medium was externally supplied and recycled at the ratio of 0.1. Therefore, acetification was conducted in both the bioreactor and reservoir resulting in an increased ETA (6 ± 0.2 g/L/day). This also aligned with the highest system AAB biomass (confirmed by scanning electron microscopy). Under the recycled oxygenated medium supply consistently high biotransformation yields (average 77.3%) were observed over nine sequential cycles. Meanwhile, an average ETA of 6.3 ± 0.2 g/L/day was obtained. This method can have practical applications in improving the efficiency and speeding up small-scale vinegar production.

Repeated cultures of Saccharomyces cerevisiae SC90 to tolerate inhibitors generated during cassava processing waste hydrolysis for bioethanol production.

Large amount of cassava pulp is produced as by-product of industrial tapioca production. The value-added process of this low-cost waste is to use it as a substrate for bioethanol production. However, during the pulp pretreatment by acidification combined with steam explosion, many yeast inhibitors including acetic acid, formic acid, levulinic acid, furfural and 5-hydroxymethylfurfural are generated and these compounds have negative effects on the subsequent fermentation step. Therefore, the objective of this study was to investigate whether the repeated cultures of Saccharomyces cerevisiae SC90 could alleviate this problem. To obtain the inhibitor tolerable cells, the repeated culture was performed by growing yeast cells to a specific growth rate (µ) of 0.22 h-1 or higher (80% of the µ in control) and then transferring them to progressively higher concentrations of hydrolysate ranging from 20 to 100% (v/v). The results showed a tendency of longer lag phase as well as time to reach maximum cell number (t maxc) with an increase in hydrolysate concentration. However, the repeated culture at the same hydrolysate concentration could shorten both lag period and t maxc. Interestingly, the growth and fermentation efficiency of adapted cells in 100% hydrolysate were significantly higher (p ≤ 0.05) than those of non-adapted cells by 38% and 27%, respectively.

Susceptibility of Klebsiella pneumoniae on coriander leaves to liquid- and vapor-phase ethanol.

The bio-control of ethanol on Klebsiella pneumoniae on fresh coriander leaves for significantly reducing consumer health risk was investigated. Washed and sterilized leaves of coriander were inoculated with K. pneumoniae cultured in Trypticase Soy broth. Susceptibility of the K. pneumoniae to liquid- and evaporated vapor-phase ethanol (EVE) was then examined in vitro Complete inhibition of K. pneumoniae was found with 18% (v/v) liquid ethanol. Exposure for 15 min to EVE (9.00 ± 0.8 mmol L(-1)) completely destroyed K. pneumoniae (4.04 ± 0.02 log CFU/ml) spread on Mueller Hilton agar at 30 ± 2°C. The effect of EVE with and without evaporated water vapor (EWV) on the susceptibility of K. pneumoniae on fresh coriander leaves was examined. While exposure to EVE affected the survival of K. pneumoniae, the degree of reduction depended on both the inoculation level and the EWV. Complete reduction of K. pneumoniae was achieved for the low inoculation level by EVE alone (37 ± 2% relative humidity; RH) but susceptibility was reduced with EWV (high RH; 80 ± 2%). Scanning electron microscope (SEM) images of inoculated coriander leaves confirm the effects of EVE in reducing levels of K. pneumoniae Exposure to EVE alone proved an effective bio-control for K. pneumoniae on fresh coriander leaves.