The removal of antibiotics and resistance genes in wastewater treatment plants has attracted widespread attention, but the potential role of residual antibiotics in the disposal of waste activated sludge (WAS) has not been clearly understood. In this study, the effect of roxithromycin (ROX) on volatile fatty acid (VFA) recovery from WAS anaerobic fermentation was investigated. The experimental results showed that ROX made a positive contribution to the production of VFAs. With the increase of ROX dosages from 0 to 100 mg/kg TSS, the maximum accumulation of VFAs increased from 295 to 610 mg COD/L. Mechanism studies revealed that ROX promoted the solubilization of WAS by facilitating the disruption of extracellular polymeric substances. In addition, ROX enhanced the activity of acetate kinase and inhibited the activities of α-glucosidase and coenzyme F420, and showed a stronger inhibitory effect on methane production than the hydrolysis process, thus resulting in an increase in VFA accumulation. These findings provide a new insight for the role of antibiotics in anaerobic fermentation of WAS.
The manifestation of class D β-lactamases in the community raises significant concern as they can hydrolyze carbapenem antibiotics. Hence, it is exceptionally alluring to design novel inhibitors. Structure-based virtual screening using docking programs and molecular dynamics simulations was employed to identify two novel non-β-lactam compounds that possess the ability to block different OXA variants. Furthermore, the presence of a nonpolar aliphatic amino acid, valine, near the active site serine, was identified in all OXA variants that can be accounted to block the catalytic activity of OXA enzymes.
β-glucosidases (Bgl) are widely utilized for releasing non-reducing terminal glucosyl residues. Nevertheless, feedback inhibition by glucose end product has limited its application. A noticeable exception has been found for β-glucosidases of the glycoside hydrolase (GH) family 1, which exhibit tolerance and even stimulation by glucose. In this study, using local isolate Trichoderma asperellum UPM1, the gene encoding β-glucosidase from GH family 1, hereafter designated as TaBgl2, was isolated and characterized via in-silico analyses. A comparison of enzyme activity was subsequently made by heterologous expression in Escherichia coli BL21(DE3). The presence of N-terminal signature, cis-peptide bonds, conserved active site motifs, non-proline cis peptide bonds, substrate binding, and a lone conserved stabilizing tryptophan (W) residue confirms the identity of Trichoderma sp. GH family 1 β-glucosidase isolated. Glucose tolerance was suggested by the presence of 14 of 22 known consensus residues, along with corresponding residues L167 and P172, crucial in the retention of the active site's narrow cavity. Retention of 40% of relative hydrolytic activity on ρ-nitrophenyl-β-D-glucopyranoside (ρNPG) in a concentration of 0.2 M glucose was comparable to that of GH family 1 β-glucosidase (Cel1A) from Trichoderma reesei. This research thus underlines the potential in the prediction of enzymatic function, and of industrial importance, glucose tolerance of family 1 β-glucosidases following relevant in-silico analyses.
Catalytic ionic liquid hydrolysis of cellulosic material have been considered as a green and highly efficient dissolution process. However, application of a pre-treatment process, i.e; ultrasonication enhances the hydrolysis of cellulose in ionic liquid by providing mechanical force. In this paper, we describe the impact of both chemical and mechanical approaches to produce nanocrytalline cellulose (NCC) with anticipated particle size, and crystallinity with improved yields. The ultrasonication treatment was evaluated in terms of treatment time and vibration amplitude. It was found that the lowest ultrasonication time (5 min) produced the NCC of highest crystallinity (73 %), but the lowest yield (84 %). In contrary, the highest ultrasonication vibration amplitude at 90 % produced NCC with highest crystallinity value (67 %) as well as yields (90 %). It concludes that ultrasonic pre-treatment improves the hydrolysis process of cellulose in ionic liquid with increasing yield and crystallinity of NCC.
Kinetic studies of the enzymatic hydrolysis of molasses were conducted using glucoamylase. Central Sugar Refinery SDN BHD contains 13-20% glucose. The molasses was diluted and the kinetic experiments were conducted at 67 degrees C with 100-1000 mg/l of glucoamylase. The glucose contents of the molasses were enhanced after hydrolysis of molasses solution with 1000 mg/l glucoamylase. A Lineweaver-Burk plot was obtained based on enzyme kinetic data. The rate constant, Km and maximum reaction rate, Vmax for 500 mg/l of glucoamylase were 100 mmol/l (18 g/l) and 5 mmol/l min (0.9 g/l min), respectively. The maximum reaction rate, Vmax for 1000 mg/l of glucoamylase was doubled, to 100 mmol/l (18 g/l) and the rate constant, Km was the same for 500 mg/l of glucoamylase. The substrate inhibition model was noncompetitive based on the resulting Lineweaver-Burk plot for enzyme concentration of 500 and 1000 mg/l.
The objective of this study is to establish conditions that allow optimal yield and antioxidant
activity for Golden Apple Snail (GAS) (Pomacea canaliculata) protein hydrolysate by employing
response surface methodology (RSM). A three level, face-centered, central composite design
(CCD) was adapted to assess the effects of temperature (45–65˚C); pH (8–10); the ratio of
enzyme to substrate (2–4%); and hydrolysis time (60–180 min). The antioxidative activity
of the hydrolysate obtained under optimized conditions was then evaluated via the following
metrics: hydroxyl radical scavenging, reducing power, and chelating effects on ferrous ion.
Established optimal conditions for the enzymatic protein hydrolysis of GAS were a temperature
of 45˚C, a pH of 10, an enzyme concentration of 2%, and hydrolysis time of 159 minutes. The
optimized GAS protein hydrolysate produced an experimental yield of 9.72% and antioxidant
activity of 73.54%—slightly less than the predicted yield of 11.36% and antioxidant activity of
78.88%. The optimized GAS protein hydrolysate formed demonstrated both higher chelating
effects and hydroxyl scavenging activity but had lower reducing power. These results suggest
that GAS protein hydrolysate holds potential as a natural antioxidant for use in food processing.
Cockle (Anadara granosa) meat wash water precipitate was hydrolyzed using bromelain. Experiments were carried out to determine optimum conditions for temperature, enzyme concentration and hydrolysis time using response surface methodology (RSM) based on a central composite rotatable design (CCRD) to obtain the highest value of nitrogen content (NC) and degree of hydrolysis (DH). Results revealed that the optimum conditions for temperature, enzyme concentration and hydrolysis time were 33.7°C, 1.45% (E/S) and 28.42 hrs, respectively. At the optimum condition, hydrolysis of cockle meat wash water precipitate using bromelain resulted in a NC of 0.6% and DH of 48%. The NC and DH were significantly influenced by temperature, enzyme concentration and hydrolysis time. When the bromelain concentration, hydrolysis time and temperature were increased, the values of NC and DH also increased. The hydrolysate produced contained flavor compounds found in clam and oyster which were 3-methylbutanol and 1-pentanol. The compound 3-MCPD was not found in the hydrolysate.
Marine invertebrates, such as oysters, mussels, clams, scallop, jellyfishes, squids, prawns, sea cucumbers and sea squirts, are consumed as foods. These edible marine invertebrates are sources of potent bioactive peptides. The last two decades have seen a surge of interest in the discovery of antioxidant peptides from edible marine invertebrates. Enzymatic hydrolysis is an efficient strategy commonly used for releasing antioxidant peptides from food proteins. A growing number of antioxidant peptide sequences have been identified from the enzymatic hydrolysates of edible marine invertebrates. Antioxidant peptides have potential applications in food, pharmaceuticals and cosmetics. In this review, we first give a brief overview of the current state of progress of antioxidant peptide research, with special attention to marine antioxidant peptides. We then focus on 22 investigations which identified 32 antioxidant peptides from enzymatic hydrolysates of edible marine invertebrates. Strategies adopted by various research groups in the purification and identification of the antioxidant peptides will be summarized. Structural characteristic of the peptide sequences in relation to their antioxidant activities will be reviewed. Potential applications of the peptide sequences and future research prospects will also be discussed.
Agriculture residues are a promising feedstock for value-added products from lignocellulosic waste. However, pretreatment of lignocellulosic materials is essential to facilitate enzymatic
hydrolysis and improve sugar yield. The objective of this study is to evaluate the effect of acid or alkali during microwave-assisted pretreatment of dragon fruit foliage (DFF) that
would make hydrolysis process more efficient. In the present study, distilled water and three chemicals were examined for their effects on releasing monomeric sugar during microwave
treatment. Microwave-assisted pretreatment namely microwave-distilled water (M-H2O) (control); microwave-sulfuric acid (M-H2SO4); microwave-sodium hydroxide (M-NaOH); and
microwave-sodium bicarbonate (M-NaHCO3) pretreatment were performed using 5% (w/v) of DFF as substrate at 800 watt microwave power for 5 minutes exposure time. Highest yield
of monomeric sugar was found at 15.56 mg/g using M-NaOH pretreatment at 0.1N NaOH. For M-H2SO4 pretreatment, 0.1N H2SO4 produced 8.2 mg/g of monomeric sugar. Application
of M-NaHCO3 pretreatment using 0.05N NaHCO3 solution released 6.45 mg/g of monomeric sugar. While, soaking DFF in distilled water and subjecting to microwave irradiation released
6.6 mg/g of monomeric sugar. Treatments with the lowest concentration (0.01 N) of the three chemicals released only small quantities of total monomeric sugars and less than that with distilled water. The changes in the physical structure of DFF prior to and after the microwaveassisted pretreatment are also reported.
The native sago starch exists as a compact crystalline structure and is not efficiently hydrolyzed by Raw Starch Degrading Enzyme (RSDE). In order to enhance its hydrolysability, the starch was treated with acid and heated below its gelatinization temperature, thus increasing the accessibility of the sago starch granule to enzymatic attack. Results showed that treatment of sago starch with acid at pH 2.0 and temperature 65oC for 2 hours greatly enhanced its conversion rate to glucose from 53.3% to 71.9%. It is clearly shown that high yield of glucose is produced during hydrolysis of acid-treated sago starch using the Raw Starch Degrading Enzyme from Acremonium sp. The difference between the acid-treated and untreated sago starch in this study could be due to the differences on the surface of the sago starch granule which may influence the accessibility and diffusion of enzyme into the starch during hydrolysis.
Utilization of palm kernel expeller (PKE), a palm oil milling by-product, may be diversified through the exploitation of its protein component. The PKE protein could be effectively extracted using an alkaline
solution and followed by enzymatic hydrolysis to produce PKE protein hydrolysates or crude PKE peptide. The extraction of PKE protein was successfully carried out using an alkaline solution at pH11, at ratio of 1:10 (g/ml), PKE powder to alkaline solution with continuous shaking, 150 rpm, in a water bath operating at 50°C for 30 min. The extracted protein powder (PKEP) had 68.50±3.08% crude protein, 0.54±0.03% fat and 0.73±0.02% ash. The freeze-dried PKEP was re-suspend in particular buffer and hydrolyzed with proteolytic enzymes (Alcalase® 2.4L, Flavourzyme® 500MG, pepsin or trypsin) to obtain PKEP hydrolysate (PKEPH). The effect of enzyme concentration (0, 2, 4, 6, 8 & 10%) and time of hydrolysis (0, 6, 12, 24, 48 h) was studied to determine the most efficient hydrolytic conditions. Results showed that all enzymes tested were capable of hydrolyzing the PKEP and producing hydrolysates with different degree of hydrolysis (DH%). At 8.0% concentration, Alcalase®2.4L hydrolyzed PKEP into the highest DH (75.96%) hydrolysate (PKEPH) after 1h hydrolysis. Although only with 2.0% Alcalase 2.4 L concentration, it was sufficient to produce PKEP hydrolysate of 81.35% DH %, but it required 12 h to hydrolyze the protein. Pepsin was relatively the least efficient protease to hydrolyze the PKEP.
Protein-rich by-products from the canning industry, especially dark flesh of skipjack, have limited uses due to several factors such as darken color, susceptibility to oxidation and off flavour. Protein hydrolysates from skipjack dark flesh was produced with different type of industrial proteases (Alcalase®2.4L FG, Protamex®, Neutrase®1.5MG and Flavourzyme®500MG) for 60, 120, 180 and 240 min with level of proteases used of 0.5, 1, 1.5 and 2% per weight of raw material. The degree of hydrolysis and free tryptophan content of hydrolysate were investigated. The results shows longer time with higher concentration of enzyme has increased the degree of hydrolysis. Alcalase®2.4L FG had the highest degree of hydrolysis among all proteases followed by Protamex®, Flavourzyme®500MG and Neutrase® 1.5MG. All enzymes increase free tryptophan content linearly with the increament of protease enzyme level. The longer the hydrolysis time, the higher the content of free tryptophan produced.
Konjac glucomannan (KGM) was treated with or without sonication and/or hydrochloric acid (HCl).
Hydrolysis effects on KGM were studied for properties such as degree of hydrolysis, fluidity, molecular mass, and creep properties. The degree of hydrolysis for sonicated KGM and acid treated KGM were not significantly different. However, the combination treatment of acid hydrolysis and sonication was found effective in yielding a smaller molecular weight fraction of KGM and solution with higher fluidity. From the creep analysis, KGM treated with combination treatment exhibited the highest compliance among samples tested. In general, ultrasound mediated acid hydrolysis was found to be a promising technique in degrading high molecular weight biopolymer. This could be attributed to a localized high temperature and high shear forces generated during cavitation that facilitated the endothermic acid hydrolysis.
The effect of degree of hydrolysis (DH) on the physicochemical properties of cobia frame hydrolysate was determined. Three levels of degree of hydrolysis of cobia frame hydrolysate were studied, which were 53%, 71% and 96%. After enzymatic hydrolysis using Alcalase®, the samples were spray-dried. Cobia hydrolysate powder samples were analyzed for their proximate analysis and physicochemical properties. The proximate analysis showed significant differences in fat and ash content only. DH96 hydrolysate showed desirable essential amino acid profile for human requirement except for methionine and isoleucine. The study found that cobia frame hydrolysate had good colour, emulsifying capacity and excellent foaming properties. However, there were no significant differences in water-holding capacity, oil-holding capacity and peptide solubility among the hydrolysate samples. This study suggested that cobia frame hydrolysate is a potential ingredient and foaming agent for food industry.
In this study, Hydrolysate from angelwing clam (Pholas orientalis) was produced at 0, 1, 2 and 3 hrs and E/S ratio of 0.5 and 3% using alcalase where the pH and temperature were kept constant at pH 8.5 and 60°C, respectively. The hydrolysates were analysed for antioxidant and functional properties such as solubility, emulsifying properties and water and oil holding capacity. Degree of hydrolysis (DH), yield, functional and antioxidant properties were influenced by the hydrolysis time and E/S ratio. Higher enzyme concentration (E/S 3%) and longer hydrolysis time increased the DH. Yield was higher at E/S 3% but reduced with hydrolysis time. Longer hydrolysis time produced more soluble hydrolysate and higher metal chelating activity but lower in emulsifying properties and DPPH activity. Higher enzyme concentration resulted in increase only in solubility and metal chelating activity. This study revealed that enzymatic hydrolysis using alcalase should be performed at shorter hydrolysis time using intermediate concentration of enzyme (E/S between 0.5 to 3%) in order to produce angelwing clam hydrolysate with collectively good functional and antioxidant properties
Banana pulp (BP) noodles prepared by partial substitution of wheat flour with green Cavendish banana pulp flour were assessed pH, color, tensile strength and elasticity, and in-vitro hydrolysis index (HI) and estimated glycemic index (GI). BP noodles had lower L* (darker) and b* values (less yellow) but higher tensile strength and elasticity modulus than control noodles. Following an in-vitro starch hydrolysis studies, it was found that GI of BP noodles was lower than control noodles. Partial substitution of green banana pulp into noodles may be useful for controlling starch hydrolysis of yellow noodles.
This study aims to determine the combined effects of hydrolysis time, temperature, pH and ratio of enzyme to substrate on the degree of hydrolysis (DH) of silver catfish frame using Response Surface Methodology. The proximate compositions of silver catfish frame and silver catfish hydrolysate powder were determined as well. The effects of independent factors were described using a three-level factors Face Centered Central Composite design. The suggested hydrolysis conditions for obtaining the optimum DH using Alcalase® were – temperature of 55oC, hydrolysis time of 163 min, pH of substrate at 9.45 and an enzyme concentration of 2.0%. The generated model showed a quadratic fit with experimental data. Proximate analyses revealed that silver catfish frame contained 25.02% protein, 68.21% fat and 7.08% ash. While silver catfish frame hydrolysate powder contained 65.05% protein, 32.92% fat and 0.86% ash. The protein recovery in silver catfish frame hydrolysate was as high as 71.6%.
In Malaysia, the amount of food waste produced is estimated at approximately 70% of total municipal solid waste generated and characterised by high amount of carbohydrate polymers such as starch, cellulose, and sugars. Considering the beneficial organic fraction contained, its utilization as an alternative substrate specifically for bioethanol production has receiving more attention. However, the sustainable production of bioethanol from food waste is linked to the efficient pretreatment needed for higher production of fermentable sugar prior to fermentation. In this work, a modified sequential acid-enzymatic hydrolysis process has been developed to produce high concentration of fermentable sugars; glucose, sucrose, fructose and maltose. The process started with hydrothermal and dilute acid pretreatment by hydrochloric acid (HCl) and sulphuric acid (H2SO4) which aim to degrade larger molecules of polysaccharide before accessible for further steps of enzymatic hydrolysis by glucoamylase. A kinetic model is proposed to perform an optimal hydrolysis for obtaining high fermentable sugars. The results suggested that a significant increase in fermentable sugar production (2.04-folds) with conversion efficiency of 86.8% was observed via sequential acid-enzymatic pretreatment as compared to dilute acid pretreatment (∼42.4% conversion efficiency). The bioethanol production by Saccharomyces cerevisiae utilizing fermentable sugar obtained shows ethanol yield of 0.42g/g with conversion efficiency of 85.38% based on the theoretical yield was achieved. The finding indicates that food waste can be considered as a promising substrate for bioethanol production.
Many researchers have focused chitosan as a source of potential bioactive material during the past few decades. However, chitosan has several drawbacks to be utilised in biological applications, including poor solubility under physiological conditions. Therefore, a new interest has recently emerged on partially hydrolysed chitosan, chitosan oligosaccharides (COS). In this study, degradation of chitosan was performed by Cellulase from Trichoderma reesei® 1.5L and Response Surface Methodology (RSM) were employed to optimize the hydrolysis temperature, pH, enzyme concentration and substrate concentration. Optimization of cellulase T. reesei® using central composite design (CCD) was to obtain optimum parameters and all the factors showed significant effects (p˂0.05). The maximum response, Celluclast® activity (1.268 U) was obtained by assaying the process at 49.79oC, pH 4.5, 3% (v/w) of enzyme concentration and 25% (w/v) concentration of chitosan for 24 hours.
Poly(lactic acid) (PLA), a bio-based polyester, has been extensively investigated in the recent past owing to its excellent mechanical properties. Several studies have been conducted on PLA blends, with a focus on improving the brittleness of PLA to ensure its suitability for various applications. However, the increasing use of PLA has increased the contamination of PLA-based products in the environment because PLA remains intact even after three years at sea or in soil. This review focuses on analyzing studies that have worked on improving the degradation properties of PLA blends and studies how other additives affect degradation by considering different degradation media. Factors affecting the degradation properties, such as surface morphology, water uptake, and crystallinity of PLA blends, are highlighted. In natural, biotic, and abiotic media, water uptake plays a crucial role in determining biodegradation rates. Immiscible blends of PLA with other polymer matrices cause phase separation, increasing the water absorption. The susceptibility of PLA to hydrolytic and enzymatic degradation is high in the amorphous region because it can be easily penetrated by water. It is essential to study the morphology, water absorption, and structural properties of PLA blends to predict the biodegradation properties of PLA in the blends.