Biological Conversion of Agricultural Wastes into Indole-3-acetic Acid by Streptomyces lavenduligriseus BS50-1 Using a Response Surface Methodology (RSM).

Agricultural waste is an alternative source for plant growth regulator biosynthesis by microorganisms. Actinobacteria are important soil microbes that significantly impact the soil as plant growth-promoting rhizobacteria and biofertilizers. This study focused on developing low-cost medium based on bagasse to improve indole-3-acetic acid (IAA) production by Streptomyces lavenduligriseus BS50-1 using a response surface methodology (RSM). Among 34 actinobacterial strains, S. lavenduligriseus BS50-1 produced the highest IAA level within the selected medium. An RSM based on a central composite design optimized the appropriate nutrients for IAA production. Thus, glucose hydrolysate and l-tryptophan at concentrations of 3.55 and 5.0 g/L, respectively, were the optimal factors that improved IAA production from 37.50 to 159.47 µg/mL within 168 h. This study reported a potential application of leftover bagasse as the raw material for cultivating actinobacteria, which efficiently produce IAA to promote plant growth.

Assessing Fermentation Broth Quality of Pineapple Vinegar Production with a Near-Infrared Fiber-Optic Probe Coupled with Stability Competitive Adaptive Reweighted Sampling.

In this study, the performance of a near-infrared (NIR) fiber-optic probe coupled with stability competitive adaptive reweighted sampling (SCARS) was investigated for the analysis of acetic acid, ethanol, total soluble solids, caffeic acid, gallic acid, and tannic acid in the broth of pineapple vinegar during fermentation. The NIR spectra of the broth samples in the region of 11,536-3956 cm-1 were collected during vinegar fermentation promoted by Acetobacter aceti. This continuous biological process led to changes in the concentrations of all analytes studied. SCARS provided optimized and stabilized NIR spectral variables for the construction of a partial least squares (PLS) model for each analyte using a small number of optimal variables (under 88 variables). The SCARS-PLS model outperformed the conventional PLS model, and achieved excellent accuracy in accordance with ISO 12099:2017 for the four prediction models of acetic acid, ethanol, caffeic acid, and gallic acid, with root-mean-square error of prediction values of 0.137%, 0.178%, 0.637 µg/mL and 0.640 µg/mL, respectively. In contrast, only an acetic acid content prediction model constructed via the conventional PLS method and using the whole spectral region (949 variables) could pass with acceptable accuracy. These results indicate that the NIR optical probe coupled with SCARS is an appropriate method for the continuous monitoring of multianalytes during vinegar fermentation, particularly acetic acid and ethanol contents, which are indicators of the finished fermentation of pineapple vinegar.

Statistical optimization of P(3HB-co-3HHx) copolymers production by Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp and its properties characterization.

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] is a bacterial copolymer in the polyhydroxyalkanoates (PHAs) family, a next-generation bioplastic. Our research team recently engineered a newly P(3HB-co-3HHx)-producing bacterial strain, Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp. This strain can produce P(3HB-co-2 mol% 3HHx) using crude palm kernel oil (CPKO) as a sole carbon substrate. However, the improvement of P(3HB-co-3HHx) copolymer production by this strain has not been studied so far. Thus, this study aims to enhance the production of P(3HB-co-3HHx) copolymers containing higher 3HHx monomer compositions using response surface methodology (RSM). Three significant factors for P(3HB-co-3HHx) copolymers production, i.e., CPKO concentration, sodium hexanoate concentration, and cultivation time, were studied in the flask scale. As a result, a maximum of 3.6 ± 0.4 g/L of P(3HB-co-3HHx) with 4 mol% 3HHx compositions was obtained using the RSM optimized condition. Likewise, the higher 3HHx monomer composition (5 mol%) was obtained when scaling up the fermentation in a 10L-stirrer bioreactor. Furthermore, the produced polymer's properties were similar to marketable P(3HB-co-3HHx), making this polymer suitable for a wide range of applications.

Biosynthesis of P(3HB-co-3HHx) Copolymers by a Newly Engineered Strain of Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp for Skin Tissue Engineering Application.

Polyhydroxyalkanoates (PHAs) are biodegradable polymers synthesized by certain bacteria and archaea with functions comparable to conventional plastics. Previously, our research group reported a newly PHA-producing bacterial strain, Rhodococcus pyridinivorans BSRT1-1, from the soil in Thailand. However, this strain's PHA synthase (phaCRp) gene has not yet been characterized. Thus, this study aims to synthesize PHA using a newly engineered bacterial strain, Cupriavidus necator PHB-4/pBBR_CnPro-phaCRp, which harbors the phaCRp from strain BSRT1-1, and characterize the properties of PHA for skin tissue engineering application. To the best of our knowledge, this is the first study on the characterization of the PhaC from R. pyridinivorans species. The results demonstrated that the expression of the phaCRp in C. necator PHB-4 had developed in PHA production up to 3.1 ± 0.3 g/L when using 10 g/L of crude palm kernel oil (CPKO) as a sole carbon source. Interestingly, the engineered strain produced a 3-hydroxybutyrate (3HB) with 2 mol% of 3-hydroxyhexanoate (3HHx) monomer without adding precursor substrates. In addition, the 70 L stirrer bioreactor improved P(3HB-co-2 mol% 3HHx) yield 1.4-fold over the flask scale without altering monomer composition. Furthermore, the characterization of copolymer properties showed that this copolymer is promising for skin tissue engineering applications.

Mushroom ß-Glucan Recovered from Antler-Type Fruiting Body of Ganoderma lucidum by Enzymatic Process and Its Potential Biological Activities for Cosmeceutical Applications.

Mushrooms are incredibly valuable macro fungi that are an important and integral part of the ecosystem. In addition to being used as cuisine, mushrooms have been used for medicinal purposes for many centuries. This research applied a process for recovering ß-glucan (BG) from the antler-type fruiting body of Ganoderma lucidum as well as tested the biological activities related to cosmeceutical applications. The characterization of complex structure was performed by fourier-transform infrared (FTIR) and nuclear magnetic resonance (MNR) spectroscopies. The obtained extract contained 40.57% BG and 7.47% protein, with the detectable bioactivities of anti-tyrosinase and antioxidation. Consequently, it showed the activity that can be used to whiten the skin by reducing or inhibiting the process of skin pigmentation. The BG also showed moderate activities of anti-collagenase, anti-elastase, and anti-hyaluronidase. The test by the HET-CAM confirmed no skin irritation of the complex extract. Based on human peripheral blood mononuclear cell (PBMC) test, the BG had no significant inhibiting effect on cell viability. In addition, the obtained BG had functional properties higher than commercially available BG, especially oil-binding capacity. These findings provided new insights into the potential application of G. lucidum BG as a polymeric material in the cosmeceutical industries.

Functionality of Yeast ß-Glucan Recovered from Kluyveromyces marxianus by Alkaline and Enzymatic Processes.

ß-Glucan (BG), one of the most abundant polysaccharides containing glucose monomers linked by ß-glycosidic linkages, is prevalent in yeast biomass that needs to be recovered to obtain this valuable polymer. This study aimed to apply alkaline and enzymatic processes for the recovery of BG from the yeast strain Kluyveromyces marxianus TISTR 5925. For this purpose, the yeast was cultivated to produce the maximum yield of raw material (yeast cells). The effective recovery of BG was then established using either an alkaline or an enzymatic process. BG recovery of 35.45% was obtained by using 1 M NaOH at 90 °C for 1 h, and of 81.15% from 1% (w/v) hydrolytic protease enzyme at 55 °C for 5 h. However, BG recovered by the alkaline process was purer than that obtained by the enzymatic process. Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy confirmed the purity, the functional groups, and the linkages of BG obtained from different recovery systems and different raw materials. The results of this study suggest that an alkaline process could be an effective approach for the solubilization and recovery of considerable purity of BG from the yeast cells. In addition, the obtained BG had comparable functional properties with commercially available BG. This study reveals the effectiveness of both chemical and biological recovery of BG obtained from yeast as a potential polymeric material.

Simultaneous Monitoring of the Evolution of Chemical Parameters in the Fermentation Process of Pineapple Fruit Wine Using the Liquid Probe for Near-Infrared Coupled with Chemometrics.

This study used Fourier transform-near-infrared (FT-NIR) spectroscopy equipped with the liquid probe in combination with an efficient wavelength selection method named searching combination moving window partial least squares (SCMWPLS) for the determination of ethanol, total soluble solids, total acidity, and total volatile acid contents in pineapple fruit wine fermentation using Saccharomyces cerevisiae var. burgundy. Two fermentation batches were produced, and the NIR spectral data of the calibration samples in the wavenumber range of 11,536-3952 cm-1 were obtained over ten days of the fermentation period. SCMWPLS coupled with second derivatives searched and optimized spectral intervals containing useful information for building calibration models of four parameters. All models were validated by test samples obtained from an independent fermentation batch. The SCMWPLS models showed better predictions (the lowest value of prediction error and the highest value of residual predictive deviation) with acceptable statistical results (under confidence limits) among the results achieved by using the whole region. The results of this study demonstrated that FT-NIR spectroscopy using a liquid probe coupled with SCMWPLS could select the optimized wavelength regions while reducing spectral points and increasing accuracy for simultaneously monitoring the evolution of four chemical parameters in pineapple fruit wine fermentation.

Anti-Inflammatory Effect of Pineapple Rhizome Bromelain through Downregulation of the NF-κB- and MAPKs-Signaling Pathways in Lipopolysaccharide (LPS)-Stimulated RAW264.7 Cells.

Bromelain is a mixture of proteolytic enzymes derived from pineapple (Ananas comosus) fruit and stem possessing several beneficial properties, particularly anti-inflammatory activity. However, the molecular mechanisms underlying the anti-inflammatory effects of bromelain are unclear. This study investigated the anti-inflammatory effects and inhibitory molecular mechanisms of crude and purified rhizome bromelains on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. RAW264.7 cells were pre-treated with various concentrations of crude bromelain (CB) or purified bromelain (PB), and then treated with LPS. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α were determined by Griess and ELISA assays. The expressions of inducible nitric oxide synthetase (iNOS), cyclooxygenase (COX)-2, nuclear factor kappa B (NF-κB), and mitogen-activated protein kinases (MAPKs)-signaling pathway-related proteins were examined by western blot analysis. The pre-treatment of bromelain dose-dependently reduced LPS-induced pro-inflammatory cytokines and mediators, which correlated with downregulation of iNOS and COX-2 expressions. The inhibitory potency of PB was stronger than that of CB. PB also suppressed phosphorylated NF-κB (p65), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 proteins in LPS-treated cells. PB then exhibited potent anti-inflammatory effects on LPS-induced inflammatory responses in RAW264.7 cells by inhibiting the NF-κB and MAPKs-signaling pathways.

Rapid selection of Andrographis paniculata medicinal plant materials based on major bioactive using near-infrared spectroscopy.

Abstract: The quantitative analysis of andrographolides in Andrographis paniculata plant materials is essential for pharmaceutical factories. This analysis cannot be done for all samples due to the conventional process using the extraction and HPLC methods requires a long analysis time and sample destruction. Therefore, near-infrared spectroscopy (NIRS) was employed to classify the class of A. paniculata and to determine the content of two active ingredients, andrographolide (AP1) and dehydroandrographolide (AP3) in A. paniculata, rapidly and non-destructively. One hundred twenty dried powder samples were obtained from aerial parts, branches, leaves, and branches mixed with leaves. The NIR absorption scans were collected from a broad spectral region (1000-2500 nm). Then, the scanned samples were extracted and analyzed for their AP1 and AP3 contents using an HPLC reference method. The success classification model based on AP1 level was developed using the second derivative pretreated NIR spectra of the entire wavelength region using the Partial Least Squares-Discriminant Analysis (PLS-DA) method. The NIR calibration models were developed and tested for quantitative analysis with 50 independent samples. The models were identified for the analysis of the AP1 content with excellent performance (correlation coefficient (R) = 0.98; standard error of validation (SEV) = 0.24%) and for the analysis of the AP3 content at a good level of efficiency (R = 0.93; SEV = 0.15%). This study showed that NIR spectroscopic method offers rapid analysis for the selection of A. paniculata that meets the requirement in bioactive amount. Supplementary Information: The online version contains supplementary material available at 10.1007/s11696-021-01746-0.

Enhanced polyhydroxybutyrate (PHB) production by newly isolated rare actinomycetes Rhodococcus sp. strain BSRT1-1 using response surface methodology.

Poly-ß-hydroxybutyrate (PHB) is a biodegradable polymer, synthesized as carbon and energy reserve by bacteria and archaea. To the best of our knowledge, this is the first report on PHB production by a rare actinomycete species, Rhodococcus pyridinivorans BSRT1-1. Response surface methodology (RSM) employing central composite design, was applied to enhance PHB production in a flask scale. A maximum yield of 3.6 ± 0.5 g/L in biomass and 43.1 ± 0.5 wt% of dry cell weight (DCW) of PHB were obtained when using RSM optimized medium, which was improved the production of biomass and PHB content by 2.5 and 2.3-fold, respectively. The optimized medium was applied to upscale PHB production in a 10 L stirred-tank bioreactor, maximum biomass of 5.2 ± 0.5 g/L, and PHB content of 46.8 ± 2 wt% DCW were achieved. Furthermore, the FTIR and 1H NMR results confirmed the polymer as PHB. DSC and TGA analysis results revealed the melting, glass transition, and thermal decomposition temperature of 171.8, 4.03, and 288 °C, respectively. In conclusion, RSM can be a promising technique to improve PHB production by a newly isolated strain of R. pyridinivorans BSRT1-1 and the properties of produced PHB possessed similar properties compared to commercial PHB.

Bio-synthesis of itaconic acid as an anti-crease finish for cellulosic fiber fabric.

Itaconic acid is an organic acid with a wide range of applications in the fields of polymer chemistry, pharmacy, agriculture, textile industry, etc. A bio-synthetic process of itaconic acid production in this study was carried out with Aspergillus terreus K17 having empty palm oil fruit bunches as a feedstock. Bio-synthesis of itaconic acid was compared with commercial maleic acid, itaconic acid and 1, 2, 3, 4-butanetetracarboxylic acid (BTCA) as anti-crease agents with sodium hypophosphate (SHP) as an esterification catalyst for cotton fabric finishing. The results showed that mechanical properties of cotton fabrics treated with bio-synthesized itaconic acid were better than those treated with the commercial ones whereas their whiteness index was lower. The best conditions for crease recovery were obtained from 8% w/v itaconic acid with 8% w/v SHP applied on cotton fabrics with a technique of 2-dip-2-nip, dried at 85 °C for 3 min and cured at 180 °C for 2 min. Even though the anti-crease properties of cotton fabrics treated with bio-synthesized itaconic acid were still lower than those treated with commercial maleic acid and BTCA, the finished cotton fibers retain the mechanical properties of cotton fabric. This study would be beneficial in producing itaconic acid as an eco-friendly anti-crease agent for cotton fabrics from waste empty palm oil fruit bunches by a bio-synthesis process.

Xylan supplement improves 1,3-propanediol fermentation by Clostridium butyricum.

Lignocellulosic biomass as co-substrate enhances the 1,3-propanediol (1,3-PD) production of anaerobic fermenters by increasing their conversion yield from glycerol. To improve 1,3-propanediol (1,3-PD) production by this efficient approach, Clostridium butyricum I5-42 was supplemented with lignocellulosic biomasses (starch free fiber (CPF) from cassava pulp and xylan) as co-substrates. The 1,3-PD production and growth of C. butyricum were considerably higher in glycerol plus CPF and xylan than in glycerol alone, whereas another major polysaccharide (cellulose co-substrate) failed to improve the 1,3-PD production. C. butyricum I5-42 showed no degradation ability on cellulose powder, and only weak activity and slight growth on xylan. However CPF supplemented with xylan strongly enhanced the transcription levels of the major enzymes of 1,3-PD production (glycerol dehydratase, 1,3-propanediol dehydrogenase, and glycerol dehydrogenase). The intracellular redox reactions maintained equal balance in the supplemented media, suggesting that CPF plus xylan promotes 1,3-PD production in the reductive pathway. This promotion is probably mediated by NADH, which is effectively regenerated by small amounts of released oligosaccharides and subsequent activation of the glycerol oxidative pathway. Both supplements also improved the 1,3-PD production at high glycerol concentration. Therefore, supplementation with lignocellulolytic polysaccharides such as xylan can improve the production and productivity of 1,3-PD from glycerol in C. butyricum. Direct supplementation of CPF with xylan in 1,3-PD production has not been previously reported.

Effect of cassava pulp supplement on 1,3-propanediol production by Clostridium butyricum.

To improve its 1,3-propanediol (1,3-PD) production, Clostridium butyricum was cultivated on glycerol medium supplemented with cassava pulp (CP). At small concentrations, the CP improved the 1,3-PD productivity of C. butyricum from (0.25±0.01)g/L/h (glycerol alone) to (0.43±0.02)g/L/h (glycerol+2g/L CP) after 24h fermentation.

Cellulose hydrolysis ability of a Clostridium thermocellum cellulosome containing small-size scaffolding protein CipA.

Mutant Clostridium thermocellum YM72 that produces small-size scaffolding protein CipA (ssCipA) was isolated from wild-type YM4. Sequencing of ssCipA revealed that two domains, cohesin 6 and cohesin 7, were not present. Cellulosome prepared from YM72 exhibited a significant reduction of hydrolysis ability on crystalline celluloses such as Sigmacell type-20 and cellulose from Halocynthia. To investigate this influence in vitro, artificial cellulosomes were assembled as recombinant CipA (rCipA) and ssCipA (rssCipA) using native free-cellulosomal subunits. The cellulosome assembled using rssCipA showed a 1.8-fold decrease in the hydrolysis of crystalline cellulose compared with that of rCipA. However, no significant differences in the hydrolysis of carboxymethylcellulose and acid-swollen cellulose were observed. One protein band was missing from the complex that was assembled using rssCipA (confirmed by native-PAGE). The missing protein was identified as CelJ, which is a major cellulosomal subunit. This suggests that insufficient cooperation of CelJ into the cellulosome results in the significant reduction of hydrolysis toward crystalline cellulose. These results indicate that cohesin 6 and 7 may be responsible for the cooperation of CelJ through cohesin and dockerin interactions, and adequate cooperation of CelJ into the cellulosome is important for significant hydrolysis of crystalline cellulose.

Growth inhibition of thermotolerant yeast, Kluyveromyces marxianus, in hydrolysates from cassava pulp.

In this study, we report the inhibition of Kluyveromyces marxianus TISTR5925 growth and ethanol fermentation in the presence of furan derivatives and weak acids (acetic acid and lactic acid) at high temperatures. Cassava pulp, obtained as the waste from starch processing, was collected from 14 starch factories located in several provinces of Thailand. At a high temperature (42 °C), the cassava pulp hydrolysate from some starch factories strongly inhibited growth and ethanol production of both K. marxianus (strain TISTR5925) and Saccharomyces cerevisiae (strain K3). HPLC detected high levels of lactic acid and acetic acid in the hydrolysates, suggesting that these weak acids impaired the growth of K. marxianus at high temperature. We isolated Trp-requiring mutants that had reduced tolerance to acetic acid compared to the wild-type. This sensitivity to acetic acid was suppressed by supplementation of the medium with tryptophan.

Efficient saccharification for non-treated cassava pulp by supplementation of Clostridium thermocellum cellulosome and Thermoanaerobacter brockii ß-glucosidase.

Cassava pulp containing 60% starch and 20% cellulose is a promising renewable source for bioethanol. The starch granule was observed to tightly bind cellulose fiber. To achieve an efficient degradation for cassava pulp, saccharification tests without pre-gelatinization treatment were carried out using combination of commercial α-amylase with cellulosome from Clostridium thermocellum S14 and ß-glucosidase (rCglT) from Thermoanaerobacter brockii. The saccharification rate for cassava pulp was shown 59% of dry matter. To obtain maximum saccharification rate, glucoamylase (GA) from C. thermocellum S14 was supplemented to the combination. The result showed gradual increase in the saccharification rate to 74% (dry matter). Supplementation of GA to the combination of commercial α-amylase, cellulosome and rCglT is powerful method for efficient saccharification of cassava pulp without pretreatment.

Direct ethanol production from cassava pulp using a surface-engineered yeast strain co-displaying two amylases, two cellulases, and ß-glucosidase.

In order to develop a method for producing fuel ethanol from cassava pulp using cell surface engineering (arming) technology, an arming yeast co-displaying α-amylase (α-AM), glucoamylase, endoglucanase, cellobiohydrase, and ß-glucosidase on the surface of the yeast cells was constructed. The novel yeast strain, possessing the activities of all enzymes, was able to produce ethanol directly from soluble starch, barley ß-glucan, and acid-treated Avicel. Cassava is a major crop in Southeast Asia and used mainly for starch production. In the starch manufacturing process, large amounts of solid wastes, called cassava pulp, are produced. The major components of cassava pulp are starch (approximately 60%) and cellulose fiber (approximately 30%). We attempted simultaneous saccharification and ethanol fermentation of cassava pulp with this arming yeast. During fermentation, ethanol concentration increased as the starch and cellulose fiber substrates contained in the cassava pulp decreased. The results clearly showed that the arming yeast was able to produce ethanol directly from cassava pulp without addition of any hydrolytic enzymes.