Yee Jiun Yap

Direct recovery of cyclodextringlycosyltransferase from Bacillus cereus using aqueous two-phase flotation

Purification of cyclodextrin glycosyl transferase (CGTase) from Bacillus cereus using polyethylene glycol (PEG)-potassium phosphates aqueous two-phase flotation (ATPF) system was studied in this paper. The effects of varying PEG molecular weight, tie-line length (TLL) value, volume ratio (VR), pH value, crude concentration and gas nitrogen flotation time were investigated. The optimal condition for purification of CGTase was attained at 18.0% (w/w) PEG 8000, 7.0% (w/w) potassium phosphates, VR of 3.0, 20% (w/w) crude load at pH 7, and 80 min nitrogen flotation time at a flow rate of 5 L/min. With this optimal condition, purification factor (PFT) of 21.8 and a yield (YT) of 97.1% were attained. CGTase was successfully purified in a single downstream processing step using the ATPF.

Interfacial partitioning behaviour of bovine serum albumin in polymer-salt aqueous two-phase system

A relationship is proposed for the interfacial partitioning of protein in poly(ethylene glycol) (PEG)-phosphate aqueous two-phase system (ATPS). The relationship relates the natural logarithm of interfacial partition coefficient, ln G to the PEG concentration difference between the top and bottom phases, Δ[PEG], with the equation ln G=AΔ[PEG]+B. Results showed that this relationship provides good fits to the partition of bovine serum albumin (BSA) in ATPS which is comprised of phosphate and PEG of four different molecular weight 1450g/mol, 2000g/mol, 3350g/mol and 4000g/mol, with the tie-line length (TLL) in the range of 44-60% (w/w) at pH 7.0. The decrease of A values with the increase of PEG molecular weight indicates that the correlation between ln G and Δ[PEG] decreases with the increase in PEG molecular weight and the presence of protein-polymer hydrophobic interaction. When temperature was increased, a non-linear relationship of ln G inversely proportional to temperature was observed. The amount of proteins adsorbed at the interface increased proportionally with the amount of BSA loaded whereas the partition coefficient, K remained relatively constant. The relationship proposed could be applied to elucidate interfacial partitioning behaviour of other biomolecules in polymer-salt ATPS.

Integration Process of Fermentation and Liquid Biphasic Flotation for Lipase Separation from Burkholderia cepacia

Liquid Biphasic Flotation (LBF) is an advanced recovery method that has been effectively applied for biomolecules extraction. The objective of this investigation is to incorporate the fermentation and extraction process of lipase from Burkholderia cepacia using flotation system. Initial study was conducted to compare the performance of bacteria growth and lipase production using flotation and shaker system. From the results obtained, bacteria shows quicker growth and high lipase yield via flotation system. Integration process for lipase separation was investigated and the result showed high efficiency reaching 92.29% and yield of 95.73%. Upscaling of the flotation system exhibited consistent result with the lab-scale which are 89.53% efficiency and 93.82% yield. The combination of upstream and downstream processes in a single system enables the acceleration of product formation, improves the product yield and facilitates downstream processing. This integration system demonstrated its potential for biomolecules fermentation and separation that possibly open new opportunities for industrial production.

Analysis of Economic and Environmental Aspects of Microalgae Biorefinery for Biofuels Production: A Review

Microalgae are considered promising feedstock for the production of biofuels and other bioactive compounds, yet there are still challenges on commercial applications of microalgae‐based products. This review focuses on the economic analysis, environmental impact, and industrial potential of biofuels production from microalgae. The cost of biofuels production remains higher compared to conventional fuel sources. However, integration of biorefinery pathways with biofuels production for the recovery of value‐added products (such as antioxidants, natural dyes, cosmetics, nutritional supplements, polyunsaturated fatty acids, and so forth) could substantially reduce the production costs. It also paves the way for sustainable energy resources by significantly reducing the emissions of CO2, NOx, SOx, and heavy metals. Large‐scale biofuels production has yet to be successfully commercialized with many roadblocks ahead and heavy competition with conventional fuel feedstock as well as technological aspects. One of the prominent challenges is to develop a cost‐effective method to achieve high‐density microalgal cultivation on an industrial scale. The biofuels industry should be boosted by Governments support in the form of subsidies and incentives, for addressing the pressing climate change issues, achieving sustainability, and energy security.

SIMPLE MODELS OF WAITING LISTS

Motivated by recent empirical studies of the length distribution of hospital waiting lists, we introduce and solve a set of models that imitate the formation of waiting lists. Patients arriving in the system must choose a waiting list to join, based on its length. At the same time patients leave the lists as they get served. The model illustrates how the power-law distributions found in the empirical studies might arise, but indicates that the mechanism causing the power-laws is unlikely to be the preferential behavior of patients or their physicians.

Green technology of liquid biphasic flotation for enzyme recovery utilizing recycling surfactant and sorbitol

Liquid biphasic flotation (LBF) system has been recognized as an efficient, green, economically sustainable and biocompatible technique for biomolecules separation and purification. The main drawbacks of the conventional process of biomolecules separation are expensive production cost, utilization of phase components that are inefficiently recycled and global pollution due to high chemical consumption and wastage. In this paper, a novel approach of LBF system for lipase recovery utilizing recycling phase components comprising surfactant and xylitol was investigated. The scope of this study focuses on pollution prevention as well as clean and environmentally friendly process for enzyme extraction via LBF. The green process proposed in this study uses phase-forming components that have recovery and recycling abilities for minimal use of chemicals for enzyme extraction. This novel method utilized Triton X-100 and xylitol for lipase extraction from Burkholderia cepacia. A few parameters were optimized to obtain high lipase separation efficiency and yield. Based on the ideal conditions of LBF, the average lipase separation efficiency and yield are 86.46 and 87.49%, correspondingly. Phase components recycling were proposed in order to reduce the chemicals consumption in LBF system. Upscaling of the recycling study exhibited consistent result with the laboratory scale. It was found that 97.20 and 98.67% of Triton X-100 and xylitol were recovered after five times of recycling and that a total of 75.87% of lipase separation efficiency was obtained. Recovery and recycling of phase components in the extraction process are established as the principal green chemistry method, which yields high separation efficiency and is economically feasible on an industrial scale.

Extraction of proteins from microalgae using integrated method of sugaring-out assisted liquid biphasic flotation (LBF) and ultrasound

In this study, a simple sugaring-out supported by liquid biphasic flotation technique combined with ultrasonication was introduced for the extraction of proteins from microalgae. Sugaring-out as a phase separation method is novel and has been used in the extraction of metal ions, biomolecules and drugs. But, its functioning in protein separation from microalgae is still unknown. In this work, the feasibility of sugaring-out coupled with ultrasound for the extraction of protein was investigated. Primary studies were carried out to examine the effect of sonication on the microalgae cell as well as the separation efficiency of the integrated method. Effect of various operating parameters such as the concentration of microalgae biomass, the location of sonication probe, sonication time, ultrasonic pulse mode (includes varying ON and OFF duration of sonication), concentration of glucose, types of sugar, concentration of acetonitrile and the flow rate in the flotation system for achieving a higher separation efficiency and yield of protein were assessed. Besides, a large-scale study of the integration method was conducted to verify the consistency of the followed technique. A maximum efficiency (86.38%) and yield (93.33%) were attained at the following optimized conditions: 0.6% biomass concentration, 200 g/L of glucose concentration, 100% acetonitrile concentration with 5 min of 5 s ON/10 s OFF pulse mode and at a flow rate of 100 cc/min. The results obtained for large scale were 85.25% and 92.24% for efficiency and yield respectively. The proposed liquid biphasic flotation assisted with ultrasound for protein separation employing sugaring-out demonstrates a high production and separation efficiency and is a cost-effective solution. More importantly, this method provides the possibility of extending its application for the extraction of other important biomolecules.

Sonication and grinding pre-treatments on Gelidium amansii seaweed for the extraction and characterization of Agarose

Various pretreatments methods including sonication and grinding were performed on red seaweed Gelidium amansii for the subsequent extraction of agarose. The agarose products are usually extracted from agar powder products from seaweeds. In this study, the agarose was extracted using a direct polyethylene glycol (PEG) method without the need to first process the agar from seaweed. The agar extract was frozen then thawed and mixed directly with PEG solution to precipitate the agarose. The quality of agarose obtained was evaluated through physico-chemical properties analysis which includes spectral technique (FTIR), melting and boiling point, gel strength and sulfate content. These properties were compared with a non-pretreated sample and it was found that the addition of pretreatment steps improved the quality of agarose but gave a slightly lower yield. The gel strength of pretreated samples was much higher and the sulfate content was lower compared to non-pretreated samples. The best pretreatment method was sonication which gave gel strength of 742 g cm-2 and sulfate content of 0.63%. The extraction of agarose can be further improved with the use of different neutralizing agents. Pretreating the seaweed shows potential in improving the quality of agarose from seaweed and can be applied for future extraction of the agarose.

Extractive Bioconversion of Gamma-Cyclodextrin and Recycling of Cyclodextrin Glycosyltransferase in Liquid Biphasic System Using Thermo-Separating Polymer

An extractive bioconversion conducted on soluble starch with cyclodextrin glycosyltransferase (CGTase) enzyme in ethylene oxide-propylene oxide (EOPO)/potassium phosphates liquid biphasic system (LBS) to extract gamma-cyclodextrin (γ-CD) was examined. A range of EOPO (with potassium phosphates) molecular weights was screen to investigate the effect of the latter on the partioning efficency of CGTase and γ-CD. The results show that the optimal top phase γ-CD yield (74.4%) was reached in 35.0% (w/w) EOPO 970 and 10.0% (w/w) potassium phosphate with 2.0% (w/w) sodium chloride. A theoretical explanation for the effect of NaCl on γ-CD was also presented. After a 2 h bioconversion process, a total of 0.87 mg/mL concentration of γ-CD was produced in the EOPO/ phosphates LBS top phase. After the extraction of top phase from LBS, four continuous repetitive batches were successfully conducted with relative CGTase activity of 1.00, 0.86, 0.45, and 0.40 respectively.