Fumaric acid production from fermented oil palm empty fruit bunches using fungal isolate K20: a comparison between free and immobilized cells.

This study investigated the potential of using steam-exploded oil palm empty fruit bunches (EFB) as a renewable feedstock for producing fumaric acid (FA), a food additive widely used for flavor and preservation, through a separate hydrolysis and fermentation process using the fungal isolate K20. The efficiency of FA production by free and immobilized cells was compared. The maximum FA concentration (3.25 g/L), with 0.034 g/L/h productivity, was observed after incubation with the free cells for 96 h. Furthermore, the production was scaled up in a 3-L air-lift fermenter using oil palm EFB-derived glucose as the substrate. The FA concentration, yield, and productivity from 100 g/L initial oil palm EFB-derived glucose were 44 g/L, 0.39 g/g, and 0.41 g/L/h, respectively. The potential for scaling up the fermentation process indicates favorable results, which could have significant implications for industrial applications.

Cellulase production under solid-state fermentation by Aspergillus sp. IN5: Parameter optimization and application.

Microbial cellulases are highly versatile catalysts with significant potential in various industries, including pulp and paper, textile manufacturing, laundry, biofuel production, food and animal feed, brewing, and agriculture. Cellulases have attracted considerable attention from the scientific community owing to their broad industrial applications and the complex nature of enzymatic systems. In the present study, a novel fungal isolate of Aspergillus sp. IN5 was used to produce cellulases. We optimized each parameter, including carbon source, incubation temperature, pH, and incubation time, for maximum cellulase production using isolate IN5 under solid-state fermentation conditions. The optimized parameters for cellulase production by isolate IN5 under solid-state fermentation were as follows: substrate, soybean residue; incubation temperature, 35 °C; pH, 7.0; and incubation duration, 5 days. These conditions resulted in the highest total cellulase activity (0.26 U/g substrate), and carboxymethyl cellulase and ß-glucosidase activities of 3.32 and 196.09 U/g substrate, respectively. The obtained fungal cellulase was used for the enzymatic hydrolysis of acid- or alkali-pretreated rice straw, which served as a model substrate. Notably, compared with acid pretreatment, the pretreatment of rice straw with diluted alkali led to higher yields of reducing sugars. Maximum reducing sugar yield (286.06 ± 2.77 mg/g substrate) was obtained after 24-h incubation of diluted alkali-pretreated rice straw mixed with an enzyme loading of 15 U/g substrate. The findings of this study provide an alternative strategy for utilizing agricultural waste and an approach to efficiently produce cellulase for the degradation of lignocellulosic materials, with promising benefits for sustainable waste management.

N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production.

BACKGROUND: Among the polyhydroxyalkanoate (PHA), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)] is reported to closely resemble polypropylene and low-density polyethylene. Studies have shown that PHA synthase (PhaC) from mangrove soil (PhaCBP-M-CPF4) is an efficient PhaC for P(3HB-co-3HHx) production and N-termini of PhaCs influence its substrate specificity, dimerization, granule morphology, and molecular weights of PHA produced. This study aims to further improve PhaCBP-M-CPF4 through N-terminal truncation. RESULTS: The N-terminal truncated mutants of PhaCBP-M-CPF4 were constructed based on the information of the predicted secondary and tertiary structures using PSIPRED server and AlphaFold2 program, respectively. The N-terminal truncated PhaCBP-M-CPF4 mutants were evaluated in C. necator mutant PHB-4 based on the cell dry weight, PHA content, 3HHx molar composition, molecular weights, and granule morphology of the PHA granules. The results showed that most transformants harbouring the N-terminal truncated PhaCBP-M-CPF4 showed a reduction in PHA content and cell dry weight except for PhaCBP-M-CPF4 G8. PhaCBP-M-CPF4 G8 and A27 showed an improved weight-average molecular weight (Mw) of PHA produced due to lower expression of the truncated PhaCBP-M-CPF4. Transformants harbouring PhaCBP-M-CPF4 G8, A27, and T74 showed a reduction in the number of granules. PhaCBP-M-CPF4 G8 produced higher Mw PHA in mostly single larger PHA granules with comparable production as the full-length PhaCBP-M-CPF4. CONCLUSION: This research showed that N-terminal truncation had effects on PHA accumulation, substrate specificity, Mw, and granule morphology. This study also showed that N-terminal truncation of the amino acids that did not adopt any secondary structure can be an alternative to improve PhaCs for the production of PHA with higher Mw in mostly single larger granules.

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.

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.

Fermentation Characteristics and Aromatic Profiles of Plum Wines Produced with Hanseniaspora thailandica Zal1 and Common Wine Yeasts.

Plum has long been cultivated in northern Thailand and evolved into products having long shelf lives. In this study, plum processing was analyzed by comparing the production of plum wine using three types of yeast, Saccharomyces cerevisiae var. burgundy, Hanseniaspora thailandica Zal1, and S. cerevisiae Lalvin EC1118. EC1118 exhibited the highest alcohol content (9.31%), similar to that of burgundy (9.21%), and H. thailandica Zal1 had the lowest alcohol content (8.07%) after 14 days of fermentation. Plum wine fermented by S. cerevisiae var. burgundy had the highest total phenolic (TP) content and antioxidant activity of 469.84 ± 6.95 mg GAE/L and 304.36 ± 6.24 µg TE/g, respectively, similar to that fermented by EC1118 (418.27 ± 3.40 mg GAE/L 288.2 ± 7.9 µg TE/g). H. thailandica Zal1 exhibited the least amount of TP content and antioxidant activity; however, the volatility produced by H. thailandica Zal1 resulted in a plum wine with a distinct aroma.

Optimization of Textile Waste Blends of Cotton and PET by Enzymatic Hydrolysis with Reusable Chemical Pretreatment.

Textile waste usually ends up in landfills and causes environmental pollution. In this study, pretreatment methods for textile recycling, including autoclaving, freezing alkali/urea soaking, and alkaline pretreatment, were applied to textile waste with various cotton/polyester blending ratios. The best condition for enzymatic hydrolysis was a 60/40 textile waste blend of cotton/polyethylene terephthalate (PET) with a reusable chemical pretreatment (15% NaOH) at 121 °C for 15 min. The hydrolysis of pretreated textile waste by cellulase was optimized using response surface methodology (RSM) based on central composite design (CCD). The optimized conditions were 30 FPU/g of enzyme loading and 7% of substrate loading, which resulted in a maximum observed value of hydrolysis yield at 89.7%, corresponding to the predicted value of 87.8% after 96 h of incubation. The findings of this study suggest an optimistic solution for textile waste recycling.

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.

Streptomyces sp. AC04842: Genomic Insights and Functional Expression of Its Latex Clearing Protein Genes (lcp1 and lcp2) When Cultivated With Natural and Vulcanized Rubber as the Sole Carbon Source.

Rubber-degrading Actinobacteria have been discovered and investigated since 1985. Only recently, through the advancement of genomic sequencing and molecular techniques, genes and pathways involved in rubber degradation are being revealed; however, the complete degradation pathway remains unknown. Streptomyces sp. AC04842 (JCM 34241) was discovered by screening at a Culture Collection Centre in Sarawak for Actinomycetes forming a clear zone on natural rubber latex agar. Streptomyces is a dominant and well-studied soil bacterium playing an important role in soil ecology including carbon recycling and biodegradation. Streptomyces sp. AC04842 draft genome revealed the presence of 2 putative latex clearing protein (lcp) genes on its chromosome and is closely related to Streptomyces cellulosae. Under the Streptomyces genus, there are a total of 64 putative lcp genes deposited in the GenBank and UniProt database. Only 1 lcp gene from Streptomyces sp. K30 has been characterized. Unlike Streptomyces sp. K30 which contained 1 lcp gene on its chromosome, Streptomyces sp. AC04842 contained 2 lcp genes on its chromosome. Streptomyces sp. AC04842 lcp1 and lcp2 amino acid sequences showed 46.13 and 69.11%, respectively, similarity to lcp sequences of Streptomyces sp. K30. Most rubber degrading strains were known to harbor only 1 lcp gene, and only recently, 2-3 lcp homologs have been reported. Several studies have shown that lcp-homolog expression increased in the presence of rubber. To study the expression of lcp1 and lcp2 genes for Streptomyces sp. AC04842, the strain was incubated in different types of rubber as the sole carbon source. In general, the lcp1 gene was highly expressed, while the lcp2 gene expression was upregulated in the presence of vulcanized rubber. Mixtures of natural and vulcanized rubber did not further increase the expression of both lcp genes compared with the presence of a specific rubber type. In this study, we paved the way to the exploration of lcp homologs and their function in degrading different types of rubber.

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.

Biosynthesized Poly(3-Hydroxybutyrate) on Coated Pineapple Leaf Fiber Papers for Biodegradable Packaging Application.

This paper is aimed at investigating the usage of biosynthesized poly(3-hydroxybutyrate) (P(3-HB)) for a coating on pineapple leaf fiber paper (PLFP). For this purpose, (P(3-HB)) was produced by Rhodococcus pyridinivorans BSRT1-1, a highly potential P(3-HB) producing bacterium, with a weight-average molecular weight (Mw) of 6.07 × 10 -5 g/mol. This biosynthesized P(3-HB) at 7.5% (w/v) was then coated on PLFP through the dip-coating technique with chloroform used as a solvent. The respective coated PLFP showed that P(3-HB) could be well coated all over on the PLFP surface as confirmed by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. The brightness and mechanical properties of PLFP could be improved by coating with biosynthesized P(3-HB) in comparison to commercially available P(3-HB) and non-coated PLFP. Furthermore, coating of P(3-HB) significantly increased the water drop penetration time on the surface of PLFP and was similar to that of the commercial P(3-HB) with the same content. The results showed that all the coated PLPF samples can be degraded under the soil burial test conditions. We have demonstrated that the P(3-HB) coated PLFP paper has the ability to prevent water drop penetration and could undergo biodegradation. Taken together, the P(3-HB) coated PLFP can be applied as a promising biodegradable paper packaging.

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.