Magnesium Sulphate and β-Alanine Enhanced the Ability of Kluyveromyces marxianus Producing Bioethanol Using Oil Palm Trunk Sap

The abundance of oil palm trunk waste generated each year has encouraged research in using its sap for fermentation to produce value-added products. One of these value-added products is bioethanol production using yeast strains. In this study, the ability of Kluyveromyces marxianus ATCC 46537 to produce bioethanol using oil palm trunk sap (OPTS) was examined. The nutrients (ammonium sulphate, di-ammonium hydrogen phosphate, magnesium sulphate, b-alanine, calcium chloride and potassium dihydrogen phosphate) required to enhance production were screened and optimised. The concentrations of bioethanol and sugars were monitored with high performance liquid chromatography. The results showed that K. marxianus could attain maximum bioethanol concentration at 16 h with a higher productivity as compared to S. cerevisiae. Magnesium sulphate and b-alanine were found to increase bioethanol production. When 7.93 g/L of magnesium sulphate and 0.90 g/L of b-alanine were supplemented to OPTS, bioethanol production increased 20% with a bioethanol yield of 0.47 g/g and a productivity of 2.22 g/L.h. Therefore, minimum supplementation of OPTS with inorganic nutrients could enhance the bioethanol production of Kluyveromyces marxianus

Optimization of Monoclonal Antibodies Purification Expressed in H-192 cells using Preparative Native-Polyacrylamide Gel Electrophoresis

Monoclonal antibodies (mAbs) are unique and specific drug molecules targeting the treatment of various diseases such as arthritis, immune disorders, infectious diseases, and cancer etc. Different methods such as antibody coupled affinity chromatography, hydrophobic interaction chromatography, etc., can be applied to purify mAbs from various sources. This article provides a simple, cost effective, preparative native-polyacrylamide gel electrophoresis (n-PAGE) technique to purify mAbs expressed in H-192 cells (Hybridoma murine cell lines) against an antigen i.e. 17-alpha-hydroxyprogesterone (17-OHP), which further can have diagnostic application to detect Congenital Adrenal Hyperplasia (CAH). Furthermore, different parameters such as concentration and volume of the feedstock (medium containing antibodies), pore size of gel, height of resolving gel etc. were optimized to obtain the maximum purity and yield of mAbs

Enzymatic Conversion of Glycerol to Glyceric Acid with Immobilised Laccase in Na-Alginate Matrix

AbstractIn this study, enzymatic oxidation of glycerol was performed for the production of glyceric acid. The effectiveness of immobilised laccase in Na-Alginate matrix was also verified. Glycerol was incubated at 25oC with 30mM of 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO), which acted as a mediator, in the presence of immobilized laccase from Trametes versicolor. Glyceric acid was quantified using HPLC, while the activity of enzyme was measured using ABTS assay. The yield of laccase activity was 90%. On the other hand, glyceric acid obtained was around 7 wt% from the total product at 24h. In conclusion, enzymatic oxidation is able to produce comparable amount of glyceric acid through a more environmental friendly method

Fabricating eco-friendly binderless fiberboard from laccase-treated rubber wood fiber.

The emission of formaldehyde vapors from adhesives such as urea formaldehyde (UF) and phenol formaldehyde (PF) is a main concern for the wood composite industry. The ability of laccase enzymes to modify the fiber in pulp industries has given hope to the wood composite industry to prepare composites without using external adhesives. In the present work, rubber wood fiber was treated with different amounts of laccase enzymes at varying time intervals. Although laccase is known to cause changes to lignin, FTIR analysis of treated fiber revealed no significant difference in the chemical composition of cellulosic fiber as compared to the untreated fiber. SEM analysis exhibited a thin uniform layer of lignin deposition on the fiber surface that may be precipitated back when the reaction reaches equilibrium. XRD showed a 10% increase (maximum) in the crystallinity index of treated fibers as compared to untreated fiber. The treated fibers were dried and pressed at different platen temperatures for various time intervals without any adhesive. Mechanical properties such as MOE, MOR, and IB were done to analyze the performance of binderless fiberboard as per ASTM D1037. Binderless fiberboard of density 750 (±10) MPa that was fabricated from the fiber treated at 6 U/g enzyme for 60 min and pressed at 200 °C for 6 min showed good performance

Nutrients content of food wastes from different sources and its pre-treatment

Food wastes are the most abundant waste available in Malaysia. It is normally disposed of together with the municipal solid waste in a sanitary or non-sanitary landfill and incinerated without any recycle efforts. Based on current households’ number of 7.67 million and a food waste generation rate of 0.54 kg/day per household [1], the amount of food waste generated is about 1.51 million tons/year. Besides recovering the energy from these food wastes through anaerobic digestion, composting, pyrolysis and hydrothermal carbonization [2], the high nutrient content of food wastes can also be used as a fermentation substrate to produce value-added product. However, the composition of daily collected food waste is varied and may affect the fermentation performance and product quality. Hence, this study aims to examine the extent of variation in the nutrients content of food wastes collected from different sources and also to evaluate the effect of hydrothermal pre-treatment on the nutrients composition of food wastes. Food wastes were collected randomly from the university’s café, Indian and Chinese restaurants, morning and night markets, and also from a household. It was then sorted, weighed, crushed and blended before further characterization. Food wastes from university’s café was then hydrothermal pre-treated at 120C for 75 min at a food waste to water ratio of 1:2. Total solid and volatile solid contents and moisture content of various untreated and pre-treated food wastes were analyzed by Standard Method, while composition of lipid was determined by Bligh & Dyer method. Protein and carbohydrate contents were determined by Bradford method and Phenol-sulphuric Acid method, respectively. Results from analysis showed huge variation between food wastes from different sources, which may due to the difference in nature of the sources. After hydrothermal pre-treatment, the total solid and volatile solid contents were lower (32% and 24%, respectively) if compare to the untreated food waste. No change of lipid content was found, but the protein and carbohydrate contents in the pre-treated hydrolysate have increased 107% and 152%, respectively. The results showed that the hydrothermal pre-treated food waste can be used either as a feed medium or a supplemental medium for fermentation process. Nevertheless, further investigation for controlling the composition in food wastes is needed if a consistent nutrient contents are required

Effect of temperature and fermentation time on protease production using decapterus macarellus fish waste

The fish processing industries generate huge amounts of by-products which cause serious environmental and health problems. The environmental problems related to waste disposal can be reduced as the low cost of fish by-products contains nutrients that can enhance microbial growth and are useful for enzyme production. This research aims to investigate the effect of temperature and fermentation time on protease production from fish waste hydrolysate by using the Bacillus strain (Bacillus cereus). The pre-treatment and fish waste hydrolysate were carried out and continued with the production of protease. The effect of fermentation time was studied every 4 h for 72 h while the effect of temperature was investigated at temperatures ranging from 30 °C to 60 °C. The results showed the maximum protease production of 45.63 U/mL at 48 h of fermentation time and 44.908 ± 6.14 U/mL at a temperature of 50 °C. The current study provides insight into the effects of cultivation conditions on protease production from local fish waste industries for further optimization study to enhance protease production

Isolation and identification of acetobacter tropicalis from selected Malaysian local fruits for potential BC production

Acetobacter spp. that are commonly found on fruits, can perform oxidation processes, resulting in acetic acid production in vinegar. Besides that, Acetobacter spp. able to produce bacterial cellulose (BC), which is an essential by-product. This present study was carried out to isolate Acetobacter spp. from selected local fruits. Species verification of the bacterial isolates was performed using molecular and bioinformatic approaches. A total of six local fruits (starfruit, jackfruit, watermelon, pineapple, honeydew & banana) were subjected to seven days of fermentation in a brown sugar solution. Acetobacter spp. were isolated from the fermented medium using bromocresol green ethanol agar as the selective medium. Thirteen bacterial isolates were obtained and subjected to molecular works, including DNA extraction and PCR amplification using universal primers, targeting the 16S rRNA genes. PCR-amplified products were selected for single-pass sequencing. BLASTn analysis of the sequencing results showed three isolates (23.1%) belonging to Acetobacter tropicalis and one isolate (7.7%) representing Gluconobacter oxydans might have potential in BC production. However, the remaining nine isolates (69.2%) hit the Lactobacillus genus. Morphological observation using FESEM showed that the BC produced by all the positive bacterial isolates is similar to dried nata de coco and BC produced by Acetobacter xylinum. In addition, four similar regions of -OH stretch (3400 - 3300 cm-1), -CH stretch (2970 to 2800 cm-1), -OH bending (1620 cm-1), and -COC stretch (1100 to 1073 cm-1) are identified in the BC samples. In the future, the isolated Acetobacter and Gluconobacter strains could be further utilized for large-scale BC production in a suitable fermentation medium

Effect of temperature and fermentation time on protease production using decapterus macarellus fish waste

The fish processing industries generate huge amounts of by-products which cause serious environmental and health problems. The environmental problems related to waste disposal can be reduced as the low cost of fish by-products contains nutrients that can enhance microbial growth and are useful for enzyme production. This research aims to investigate the effect of temperature and fermentation time on protease production from fish waste hydrolysate by using the Bacillus strain (Bacillus cereus). The pre-treatment and fish waste hydrolysate were carried out and continued with the production of protease. The effect of fermentation time was studied every 4 h for 72 h while the effect of temperature was investigated at temperatures ranging from 30 °C to 60 °C. The results showed the maximum protease production of 45.63 U/mL at 48 h of fermentation time and 44.908 ± 6.14 U/mL at a temperature of 50 °C. The current study provides insight into the effects of cultivation conditions on protease production from local fish waste industries for further optimization study to enhance protease production

Size-selective purification of hepatitis B virus-like particle in flow-through chromatography: types of ion exchange adsorbent and grafted polymer architecture

Hepatitis B virus-like particles expressed in Escherichia coli were purified using anion exchange adsorbents grafted with polymer poly(oligo(ethylene glycol) methacrylate) in flow-through chromatography mode. The virus-like particles were selectively excluded, while the relatively smaller sized host cell proteins were absorbed. The exclusion of virus-like particles was governed by the accessibility of binding sites (the size of adsorbents and the charge of grafted dextran chains) as well as the architecture (branch-chain length) of the grafted polymer. The branch-chain length of grafted polymer was altered by changing the type of monomers used. The larger adsorbent (90 μm) had an approximately twofold increase in the flow-through recovery, as compared to the smaller adsorbent (30 μm). Generally, polymer-grafted adsorbents improved the exclusion of the virus-like particles. Overall, the middle branch-chain length polymer grafted on larger adsorbent showed optimal performance at 92% flow-through recovery with a purification factor of 1.53. A comparative study between the adsorbent with dextran grafts and the polymer-grafted adsorbent showed that a better exclusion of virus-like particles was achieved with the absorbent grafted with inert polymer. The grafted polymer was also shown to reduce strong interaction between binding sites and virus-like particles, which preserved the particles' structure

STUDY OF DIFFERENT TREATMENT METHODS ON CHICKEN FEATHER BIOMASS

The chicken feathers (CFs)  consist of up to 10 % of total chicken dry mass and they have many potential industrial applications. CFs contains protein fibers named as keratin, which is an insoluble protein. Primary sanitization phases are complex because of the presence of lots of blood born microbes, pathogens and parasites in raw biomass. The extraction process of keratins from the unprocessed feathers is also a challenging task. Prior to the extraction cleaning/sanitization of feathers is a very necessary step. Thus, the present work was conducted to optimize  an efficient surfactant  for the cleaning process of the  CFs by using ionic and non-ionic surfactants. The experiment was conducted by the washing of feathers with double distilled water (ddH2O), detergents, ether and lastly with boiling water. The washed feathers treated with surfactants and the effect of each surfactant was analyzed by a microbiological test which tells about the extent of  the presence of different bacteria on the treated feathers. SEM, EDX, FTIR were used to study the morphology and composition of  untreated and treated CFs. SEM showed there was no detectable fiber damage after treatment. Cetrimonium bromide (CTAB) (t3) was one of the best surfactant for the treatment of CFs among all the surfactant used. The present study described the best treatment method  for the CFs.