In this study, the ability of Cupriavidus sp. USMAA2-4 to synthesize polyhydroxyalkanoates (PHA) containing 4-hydroxyvalerate monomer (4HV) was studied through one-stage cultivation using γ-valerolactone as the carbon precursor. The presence of 4HV monomer unit in the polymer was detected through gas chromatography analysis, proving the capability of this wild strain bacterium to produce poly(3-hydrxybutyrate-co-3-hydroxyvalerate-co-4-hydroxyvalerate) [P(3HB-co-3HV-co-4HV)] terpolymer. Existence of a 4HV monomer unit in the PHA produced was further confirmed through (13)C and (1)H NMR analysis. P(3HB-co-88 % 3HV-co-1 % 4HV) terpolymer with the highest PHA content of 63 wt% was obtained through combination of 0.14 wt% C of γ-valerolactone with 0.42 wt% C of oleic acid. Various compositions of P(3HB-co-3HV-co-4HV) terpolymer with 3HV and 4HV compositions ranging from 11 to 94 mol% and from 1 to 4 mol%, respectively, were acquired by manipulating γ-valerolactone and oleic acid concentrations. The molecular weight and the thermal and mechanical properties of four different compositions of terpolymers-P(3HB-co-91 % 3HV-co-1 % 4HV), P(3HB-co-55 % 3HV-co-2 % 4HV), P(3HB-co-27 % 3HV-co-2 % 4HV), and P(3HB-co-9 % 3HV-co-1 % 4HV)-were characterized. Among these terpolymers, P(3HB-co-27 % 3HV-co-2 % 4HV) terpolymer with a molecular weight of 5.7 (10(5) Da) exhibited the highest elongation to break (264 %). The monomer unit compositional distributions of these terpolymers were investigated through acetone-water fractionation analysis. The results suggested that these produced terpolymers had broad 3HV compositional distribution and narrow 4HV compositional distribution.
Garcinia mangostana pericarp is a good source of natural antioxidants with numerous functional properties. The conventional approaches for the recovery of antioxidants from Garcinia mangostana pericarp require long processing time and high temperature, which may cause degradation or loss of bioactivity of antioxidants, and often result in low recovery efficiency. In this study, the extraction of antioxidants from Garcinia mangostana pericarp was investigated using a polyethylene glycol (PEG)/citrate aqueous biphasic system (ABS) with the addition of surfactants. The optimum condition for the recovery of antioxidants was achieved in PEG 1000/citrate ABS of pH 8 with tie-line length (TLL) of 48.3% (w/w), volume ratio (VR) of 1.6, 0.2% (w/w) sample loading and addition of 1.0% (w/w) Tween 85. The antioxidants were recovered in the PEG-rich top phase with a high K value of 18.23 ± 0.33 and a recovery yield of 92.01% ± 0.09. The findings suggested that the addition of surfactants to polymer/salt ABS can enhance the recovery of antioxidants from Garcinia mangostana pericarps by conserving the antioxidative properties.
Oil pollution in marine environment caused by oil spillage has been a main threat to the ecosystem including the ocean life and to the human being. In this research, three indigenous purple photosynthetic strains Rhodopseudomonas sp. DD4, DQ41, and FO2 were isolated from oil-contaminated coastal zones in Vietnam. The cells of these strains were immobilized on different carriers including cinder beads (CB), coconut fiber (CF), and polyurethane foam (PUF) for diesel oil removal from artificial seawater. The mixed biofilm formed by using CB, CF, and PUF as immobilization supports degraded 90, 91, and 95% of diesel oil (DO) with the initial concentration of 17.2 g/L, respectively, after 14 days of incubation. The adsorption of DO on different systems was accountable for the removal of 12-16% hydrocarbons for different carriers. To the best of our knowledge, this is the first report on diesel oil degradation by purple photosynthetic bacterial biofilms on different carriers. Moreover, using carriers attaching purple photosynthetic bacteria to remove diesel oil in large scale is considered as an essential method for the improvement of a cost-effective and efficient bioremediation manner. This study can be a promising approach to eliminate DO from oil-contaminated seawater.
Abscisic acid (ABA) has been known to exist in a microalgal system and serves as one of the chemical stimuli in various biological pathways. Nonetheless, the involvement of ABA in fatty acid biosynthesis, particularly at the transcription level in microalgae is poorly understood. The objective of this study was to determine the effects of exogenous ABA on growth, total oil content, fatty acid composition, and the expression level of beta ketoacyl-ACP synthase I (KAS I) and omega-3 fatty acid desaturase (ω-3 FAD) genes in Chlorella vulgaris UMT-M1. ABA was applied to early stationary C. vulgaris cultures at concentrations of 0, 10, 20, and 80 μM for 48 h. The results showed that ABA significantly increased biomass production and total oil content. The increment of palmitic (C16:0) and stearic (C18:0) acids was coupled by decrement in linoleic (C18:2) and α-linolenic (C18:3n3) acids. Both KAS I and ω-3 FAD gene expression were downregulated, which was negatively correlated to saturated fatty acid (SFAs), but positively correlated to polyunsaturated fatty acid (PUFA) accumulations. Further analysis of both KAS I and ω-3 FAD promoters revealed the presence of multiple ABA-responsive elements (ABREs) in addition to other phytohormone-responsive elements. However, the role of these phytohormone-responsive elements in regulating KAS I and ω-3 FAD gene expression still remains elusive. This revelation might suggest that phytohormone-responsive gene regulation in C. vulgaris and microalgae as a whole might diverge from higher plants which deserve further scientific research to elucidate its functional roles.
The oil palm sector is one of the major plantation industries in Malaysia. Palm kernel cake is a byproduct of extracted palm kernel oil. Mostly palm kernel cake is wasted or is mixed with other nutrients and used as animal feed, especially for ruminant animals. Recently, palm kernel cake has been identified as an important ingredient for the formulation of animal feed, and it is also exported especially to Europe, South Korea, and Japan. It can barely be consumed by nonruminant (monogastric) animals owing to the high percentages of hemicellulose and cellulose contents. Palm kernel cake must undergo suitable pretreatment in order to decrease the percentage of hemicellulose and cellulose. One of the methods employed in this study is fermentation with microorganisms, particularly fungi, to partially degrade the hemicellulose and cellulose content. This work focused on the production of enzymes by Aspergillus niger and profiling using palm kernel cake as carbon source.
Strategies to immobilize the individual enzymes are crucial for enhancing catalytic applicability and require a controlled immobilization process. Herein, protocol for immobilizing Candida rugosa lipase (CRL) onto modified magnetic silica derived from oil palm leaves ash (OPLA) was optimized for the effects of concentration of CRL, immobilization time, and temperature, monitored by titrimetric and spectrometric methods. XRD and TGA-DTG spectrometric observations indicated that OPLA-silica was well coated over magnetite (SiO2-MNPs) and CRLs were uniformly bound by covalent bonds to SiO2-MNPs (CRL/Gl-A-SiO2-MNPs). The optimized immobilization protocol showed that in the preparation of CRL/Gl-A-SiO2-MNPs, CRL with 68.3 mg/g protein loading and 74.6 U/g specific activity was achieved using 5 mg/mL of CRL, with an immobilization time of 12 h at 25 °C. The present work also demonstrated that acid-pretreated OPLA is a potential source of renewable silica, envisioning its applicability for practical use in enzymatic catalysis on solid support.
Neuroblastoma cell lines such as SH-SY5Y are the most frequently utilized models in neurodegenerative research, and their use has advanced the understanding of the pathology of neurodegeneration over the past few decades. In Alzheimer's disease (AD), several pathogenic mutations have been described, all of which cause elevated levels of pathological hallmarks such as amyloid-beta (Aβ). Although the genetics of Alzheimer's disease is well known, familial AD only accounts for a small number of cases in the population, with the rest being sporadic AD, which contains no known mutations. Currently, most of the in vitro models used to study AD pathogenesis only examine the level of Aβ42 as a confirmation of successful model generation and only perform comparisons between wild-type APP and single mutants of the APP gene. Recent findings have shown that the Aβ42/40 ratio in cerebrospinal fluid (CSF) is a better diagnostic indicator for AD patients than is Aβ42 alone and that more extensive Aβ formation, such as accumulation of intraneuronal Aβ, Aβ plaques, soluble oligomeric Aβ (oAβ), and insoluble fibrillar Aβ (fAβ) occurs in TgCRND8 mice expressing a double-mutant form (Swedish and Indiana) of APP, later leading to greater progressive impairment of the brain. In this study, we generated SH-SY5Y cells stably transfected separately with wild-type APP, the Swedish mutation of APP, and the Swedish and Indiana mutations of APP and evaluated the APP expression as well as the Aβ42/40 ratio in those cells. The double-mutant form of APP (Swedish/Indiana) expressed markedly high levels of APP protein and showed a high Aβ2/40 ratio compared to wild-type and single-mutant cells.
Ulcerative colitis (UC) is a serious health condition and defined as inflammation in the colon. Untreated, UC can develop into colitis-associated cancer (CAC), for which effective medicines are not available. Natural products are a better choice to treat UC by alleviating the inflammation. Caffeic acid phenethyl ester (CAPE) is a phenolic compound and known for its beneficial effects, including antibacterial, anti-inflammatory, anti-diabetic, and anticancer. We aimed to study the effect of CAPE on dextran sulfate sodium (DSS)-induced UC in mouse model. Administration of CAPE to DSS-induced mice protected against colon damage by improving body weight of mice, reducing the weight of spleen, and increased colon length. In addition, administration of CAPE resulted reduced the activity of myeloperoxidase (MPO) and CD68+ positive cells. Furthermore, a significant decrease in the production of key cytokines and the expression of nuclear factor (p65-NF)-κB. Moreover, p65-NF-κB activation was reduced in lipopolysaccharide (LPS)-treated RAW 264.7 macrophage cells from mouse origin. CAPE treatment leads to the reduced expressions of intercellular adhesion molecules (ICAM)-1 and vascular cell adhesion molecules (VCAM), both are key cell adhesion molecules. The results of this study clearly indicate that CAPE can potentially control inflammation in the colon and can be used as a therapy for UC.
The conversion of starchy sago (Metroxylon sagu) pith waste (SPW), a lignocellulosic biomass waste, to fermentable sugars under mild conditions had been successfully demonstrated. The optimum depolymerization of SPW was achieved at 2 wt% sample loading which was catalyzed by 100 mM of oxalic acid in the presence of 25 wt% NaCl solution at 110 °C for 3 h. Up to 97% SPW sample was being converted into fermentable sugars with limited formation of by-products after two sequential depolymerization cycles. Both reaction temperature and concentration of oxalic acid were crucial parameters for the depolymerization of SPW which exhibited a high selectivity for the production of glucose over other reducing sugars.
Dendrobium sonia-28 is an important ornamental orchid in the Malaysian flower industry. However, the genus faces both low germination rates and the risk of producing heterozygous progenies. Cryopreservation is currently the favoured long-term storage method for orchids with propagation problems. Vitrification, a frequently used cryopreservation technique, involves the application of pretreatments and cryoprotectants to protect and recover explants during and after storage in liquid nitrogen. However, cryopreservation may cause osmotic injuries and toxicity to cryopreserved explants from the use of highly concentrated additives, and cellular injuries from thawing, devitrification and ice formation. Reactive oxygen species (ROS), occurring during dehydration and cryopreservation, may also cause membrane damage. Plants possess efficient antioxidant systems such as the superoxide dismutase (SOD) and catalase (CAT) enzymes to scavenge ROS during low temperature stress. In this study, protocorm-like bodies (PLBs) of Dendrobium sonia-28 were assayed for the total protein content, and both SOD and CAT activities, at each stage of a vitrification exercise to observe for deleterious stages in the protocol. The results indicated that cryopreserved PLBs of Dendrobium sonia-28 underwent excessive post-thawing oxidative stress due to decreased levels of the CAT enzyme at the post-thawing recovery stage, which contributed to the poor survival rates of the cryopreserved PLBs.
Hellenia speciosa (J.Koenig) S.R. Dutta is a plant species belonging to the family Costaceae. It is widely distributed in China, India, Malaysia, Indonesia, tropical, and subtropical Asia. In Ayurveda, the rhizome of this plant has been extensively used to treat fever, rash, asthma, bronchitis, and intestinal worms. The objective of the present study was to investigate the phytochemical constituents of the leaf of Hellenia speciosa using gas chromatography and mass spectroscopy analysis (GC-MS). The GC-MS analysis revealed the presence of 17 phytochemical components in the ethanolic leaf extract of Hellenia speciosa. The prevailing bioactive compounds present in Hellenia speciosa were thymol (RT-10.019; 3.59%), caryophyllene (RT-11.854; 0.62%), caryophyllene oxide (RT-13.919; 1.34%), artumerone (RT-14.795; 1.35%), hexadecanoic acid methyl ester (RT-17.536; 2.77%), 9,12-octadecanoic acid methyl ester (RT-19.163; 1.35%), squalene (RT-24.980; 1.19%), piperine (RT-25.745; 3.11%), beta tocopherol (RT-26.681; 2.88%), vitamin E (RT-27.290; 2.64%), progesterone (RT-29.608; 3.18%), caparratriene (RT-29.861; 9.72%), and testosterone (RT-30.73; 5.81%). The compounds were identified by comparing their retention time and peak area with that of the literature and by interpretation of mass spectra. The results and findings of the present study suggest that the plant leaf can be used as a valuable source in the field of herbal drug discovery. The presence of bioactive compounds justifies the use of plant leaves for treating various diseases with fewer side effects and recommended the plant of pharmaceutical importance. However, further studies are needed to undertake its bioactivity and toxicity profile.
Xylitol production by bioconversion of xylose can be economically interesting if the raw material can be recovered from a cheap lignocellulosic biomass (LCB). Meranti wood sawdust (MWS) is a renewable and low-cost LCB that can be used as a promising and economic source of xylose, a starting raw material for the manufacture of several specialty chemicals, especially xylitol. This study aimed to optimize the hydrolysis process of MWS and to determine the influence of temperature, H2SO4 concentration, and residence time on xylose release and on by-product formation (glucose, arabinose, acetic acid, furfural, hydroxymethylfurfural (HMF), and lignin degradation products (LDPs)). Batch hydrolysis was conducted under various operating conditions, and response surface methodology was adopted to achieve the highest xylose yield. Xylose production was highly affected by temperature, acid concentration, and residence time. The optimum temperature, acid concentration, and time were determined to be 124 °C, 3.26 %, and 80 min, respectively. Under these optimum conditions, xylose yield and selectivity were attained at 90.6 % and 4.05 g/g, respectively.
Xylose reductase (XR) is an intracellular enzyme, which catalyzes xylose to xylitol conversion in the microbes. It has potential biotechnological applications in the manufacture of various commercially important specialty bioproducts including xylitol. This study aimed to prepare XR from adapted strain of Candida tropicalis and to characterize it. The XR was isolated from adapted C. tropicalis, cultivated on Meranti wood sawdust hemicellulosic hydrolysate (MWSHH)-based medium, via ultrasonication, and was characterized based on enzyme activity, stability, and kinetic parameters. It was specific to NADPH with an activity of 11.16 U/mL. The enzyme was stable at pH 5-7 and temperature of 25-40 °C for 24 h and retained above 95 % of its original activity after 4 months of storage at -80 °C. The K m of XR for xylose and NADPH were 81.78 mM and 7.29 μM while the V max for them were 178.57 and 12.5 μM/min, respectively. The high V max and low K m values of XR for xylose reflect a highly productive reaction among XR and xylose. MWSHH can be a promising xylose source for XR preparation from yeast.
Xylose-rich sawdust hydrolysate can be an economic substrate for the enzymatic production of xylitol, a specialty product. It is important to identify the process factors influencing xylitol production. This research aimed to screen the parameters significantly affecting bioxylitol synthesis from wood sawdust by xylose reductase (XR). Enzymatic bioxylitol production was conducted to estimate the effect of different variables reaction time (2-18 h), temperature (20-70 °C), pH (4.0-9.0), NADPH (1.17-5.32 g/L), and enzyme concentration (2-6 %) on the yield of xylitol. Fractional factorial design was followed to identify the key process factors. The screening design identified that time, temperature, and pH are the most significant factors influencing bioxylitol production among the variables with the values of 12 h, 35 °C, and 7.0, respectively. These conditions led to a xylitol yield of 71 % (w/w). This is the first report on the statistical screening of process variables influencing enzyme-based bioxylitol production from lignocellulosic biomass.
Synthesis of layered double hydroxides (LDHs) of Zn/Al-NO3- hydrotalcite (HIZAN) and Zn/Al-diocytyl sodium sulfosuccinate (DSS) nanocomposite (NAZAD) with a molar ratio of Zn/Al of 4:1 were carried out by coprecipitation through continuous agitation. Their structures were determined using X-ray diffractometer spectra, which showed that basal spacing for LDH synthesized by both methods was about 8.89 A. An expansion of layered structure of about 27.9 A was observed to accommodate the surfactant anion between the interlayer. This phenomenon showed that the intercalation process took place between the LDH interlayer. Lipase from Candida rugosa was immobilized onto these materials by physical adsorption method. It was found that the protein loading onto NAZAD is higher than HIZAN. The activity of immobilized lipase was investigated through esterification of oleic acid and 1-butanol in hexane. The effects of pore size, surface area, reaction temperature, thermostability of the immobilized lipases, storage stability in organic solvent, and leaching studies were investigated. Stability was found to be the highest in the nanocomposite NAZAD.
Candida rugosa lipase was modified via reductive alkylation to increase its hydrophobicity to work better in organic solvents. The free amino group of lysines was alkylated using propionaldehyde with different degrees of modification obtained (49 and 86%). Far-ultraviolet circular dichroism (CD) spectroscopy of the lipase in aqueous solvent showed that such chemical modifications at the enzyme surface caused a loss in secondary and tertiary structure that is attributed to the enzyme unfolding. Using molecular modeling, we propose that in an aqueous environment the loss in protein structure of the modified lipase is owing to disruption of stabilizing salt bridges, particularly of surface lysines. Indeed, molecular modeling and simulation of a salt bridge formed by Lys-75 to Asp-79, in a nonpolar environment, suggests the adoption of a more flexible alkylated lysine that may explain higher lipase activity in organic solvents on alkylation.
Microbial pigments are gaining intensive attention due to increasing awareness of the toxicity of synthetic colours. In this study, a novel polymer-producing bacterium designated as Cupriavidus sp. USMAHM13 was also found to produce yellow pigment when cultivated in nutrient broth. Various parameters such as temperature, pH and ratio of culture volume to flask volume were found to influence the yellow pigment production. UV-Visible, Fourier transform infrared and (13)C-nuclear magnetic resonance analyses revealed that the crude yellow pigment might probably represent new bioactive compound in the carotenoid family. The crude yellow pigment also exhibited a wide spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria with their inhibition zones and minimal inhibitory concentrations ranged from 25 to 38 mm and from 0.63 to 2.5 mg/ml, respectively. To the best of our knowledge, this is the first report on the identification and characterization of yellow pigment produced by bacterium belonging to the genus Cupriavidus.
Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) [P(3HB-co-3HV-co-4HB)] terpolymer was produced using Cupriavidus sp. USMAA2-4 via one-step cultivation process through combination of various carbon sources such as 1,4-butanediol or γ-butyrolactone with either 1-pentanol, valeric acid, or 1-propanol. Oleic acid was added to increase the biomass production. The composition of 3HV and 4HB monomers were greatly affected by the concentration of 1,4-butanediol and 1-pentanol. Terpolymers with 3HV and 4HB molar fractions ranging from 2 to 41 mol.% and 5 to 31 mol.%, respectively, were produced by varying the concentration of carbon precursors. The thermal and mechanical properties of the terpolymers containing different proportions of the constituent monomers were characterized using gel permeation chromatography (GPC), DSC, and tensile machine. GPC analysis showed that the molecular weights (M (w)) of the terpolymer produced were within the range of 346 to 1,710 kDa. The monomer compositions of 3HV and 4HB were also found to have great influences on the thermal and mechanical properties of the terpolymer P(3HB-co-3HV-co-4HB) produced.
Sacha Inchi (Plukenetia volubilis L.) is a plant native in the Amazon rainforest in South America known for its edible seeds, which are rich in lipids, proteins, vitamin E, polyphenols, minerals, and amino acids. Rural communities in developing nations have been using this plant for its health benefits, including as a topical cream for rejuvenating and revitalising skin and as a treatment for muscle pain and rheumatism. Although Sacha Inchi oil has been applied topically to soften skin, treat skin diseases, and heal wounds, its protein-rich seeds have not yet received proper attention for extensive investigation. Proteins in Sacha Inchi seeds are generally known to have antioxidant and antifungal activities and are extensively used nowadays in making protein-rich food alternatives worldwide. Notably, large-scale use of seed proteins has begun in nanoparticle and biofusion technologies related to the human health-benefitting sector. To extract and identify their proteins, the current study examined Sacha Inchi seeds collected from the Malaysian state of Kedah. Our analysis revealed a protein concentration of 73.8 ± 0.002 mg/g of freeze-dried seed flour. Employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PEAKS studio analysis, we identified 217 proteins in the seed extract, including 152 with known proteins and 65 unknown proteins. This study marks a significant step towards comprehensively investigating the protein composition of Sacha Inchi seeds and elucidating their potential applications in the food and biopharmaceutical sectors. Our discoveries not only enhance our knowledge of Sacha Inchi's nutritional characteristics but also pave the way for prospective research and innovative advancements in the realms of functional food and health-related domains.
The inexorable exposure of plants to the combinations of abiotic stresses has affected the worldwide food supply. The crop improvement against these abiotic stresses has been captivating approach to increase the yield and enhance the stress tolerance. By using traditional and modern breeding methods, the characters that confer tolerance to these stresses were accomplished. No doubt genetic engineering and molecular breeding have helped in comprehending the intricate nature of stress response. Understanding of abiotic stress-involved cellular pathways provides vital information on such responses. On the other hand, genomic research for crop improvement has raised new assessments in breeding new varieties against abiotic stresses. Interpretation of responses of the crop plants under stress is of great significance by studying the main role of crops in food and biofuel production. This review presents genomic-based approaches revealing the complex networks controlling the mechanisms of abiotic stress tolerance, and the possible modes of assimilating information attained by genomic-based approaches due to the advancement in isolation and functional analysis of genes controlling the yield and abiotic stress tolerance are discussed.