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Chemical recycling of polyhydroxyalkanoates as a method towards sustainable development

Ariffin H, Nishida H, Hassan MA, Shirai Y

Biotechnol J, 2010 May;5(5):484-92.
PMID: 20408140 DOI: 10.1002/biot.200900293

Chemical recycling of bio-based polymers polyhydroxyalkanoates (PHAs) by thermal degradation was investigated from the viewpoint of biorefinery. The thermal degradation resulted in successful transformation of PHAs into vinyl monomers using alkali earth compound (AEC) catalysts. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)s (PHBVs) were smoothly and selectively depolymerized into crotonic (CA) and 2-pentenoic (2-PA) acids at lower degradation temperatures in the presence of CaO and Mg(OH)(2) as catalysts. Obtained CA from 3-hydroxybutyrate sequences in PHBV was copolymerized with acrylic acid to produce useful water-soluble copolymers, poly(crotonic acid-co-acrylic acid) that have high glass-transition temperatures. The copolymerization of CA derived from PHA pyrolysis is an example of cascade utilization of PHAs, which meets the idea of sustainable development.
Measuring organic carbon, nutrients and heavy metals in rivers receiving leachate from controlled and uncontrolled municipal solid waste (MSW) landfills

Yusof N, Haraguchi A, Hassan MA, Othman MR, Wakisaka M, Shirai Y

Waste Manag, 2009 Oct;29(10):2666-80.
PMID: 19564103 DOI: 10.1016/j.wasman.2009.05.022

Since landfilling is the common method of waste disposal in Malaysia, river water is greatly exposed to the risk of contamination from leachate unless proper leachate management is carried out. In this study, leachates from three different types of landfills, namely active uncontrolled, active controlled and closed controlled, were characterized, and their relationships with river water chemistry were examined monthly for a year. The influence of leachate on river water chemistry from each type of landfill depended on many factors, including the presence of a leachate control mechanism, leachate characteristics, precipitation, surface runoff and the applied treatment. The impact of leachate from an active uncontrolled landfill was the highest, as the organic content, NH(4)(+)-N, Cd and Mn levels appeared high in the river. At the same time, influences of leachate were also observed from both types of controlled landfills in the form of inorganic nitrogen (NH(4)(+)-N, NO(3)(-)-N and NO(2)(-)-N) and heavy metals (Fe, Cr, Ni and Mn). Improper treatment practice led to high levels of some contaminants in the stream near the closed controlled landfill. Meanwhile, the active controlled landfill, which was located near the coastline, was exposed to the risk of contamination resulting from the pyrite oxidation of the surrounding area.
Enzyme production and profile by Aspergillus niger during solid substrate fermentation using palm kernel cake as substrate

Ong LG, Abd-Aziz S, Noraini S, Karim MI, Hassan MA

Appl Biochem Biotechnol, 2004 8 12;118(1-3):73-9.
PMID: 15304740

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.
Baseline study of methane emission from open digesting tanks of palm oil mill effluent treatment

Yacob S, Hassan MA, Shirai Y, Wakisaka M, Subash S

Chemosphere, 2005 Jun;59(11):1575-81.
PMID: 15894045

Anthropogenic release of greenhouse gases, especially CO2 and CH4 has been recognized as one of the main causes of global warming. Several measures under the Kyoto Protocol 1997 have been drawn up to reduce the greenhouse gases emission. One of the measures is Clean Development Mechanisms (CDM) that was created to enable developed countries to cooperate with developing countries in emission reduction activities. In Malaysia, palm oil industry particularly from palm oil mill effluent (POME) anaerobic treatment has been identified as an important source of CH4. However, there is no study to quantify the actual CH4 emission from the commercial scale wastewater treatment facility. Hence, this paper shall address the CH4 emission from the open digesting tanks in Felda Serting Hilir Palm Oil Mill. CH4 emission pattern was recorded for 52 weeks from 3600 m3 open digesting tanks. The findings indicated that the CH4 content was between 13.5% and 49.0% which was lower than the value of 65% reported earlier. The biogas flow rate ranged between 0.8l min(-1)m(-2) and 9.8l min(-1)m(-2). Total CH4 emission per open digesting tank was 518.9 kgday(-1). Relationships between CH4 emission and total carbon removal and POME discharged were also discussed. Fluctuation of biogas production was observed throughout the studies as a result of seasonal oil palm cropping, mill activities, variation of POME quality and quantity discharged from the mill. Thus only through long-term field measurement CH4 emission can be accurately estimated.
Soluble inhibitors generated during hydrothermal pretreatment of oil palm mesocarp fiber suppressed the catalytic activity of Acremonium cellulase

Zakaria MR, Hirata S, Fujimoto S, Ibrahim I, Hassan MA

Bioresour Technol, 2016 Jan;200:541-7.
PMID: 26524253 DOI: 10.1016/j.biortech.2015.10.075

Oil palm mesocarp fiber was subjected to hydrothermal pretreatment under isothermal and non-isothermal conditions. The pretreated slurries were separated by filtration, pretreated liquids and solids were characterized. An enzymatic digestibility study was performed for both pretreated slurries and solids to understand the effect of soluble inhibitors generated during the pretreatment process. The highest glucose yield obtained from pretreated slurries was 70.1%, and gradually decreased with higher pretreatment severities. The highest glucose yield obtained in pretreated solids was 100%, after pretreatment at 210°C for 20min. In order to study the inhibitory effects of compounds generated during pretreatment with cellulase, technical grade solutions that mimic the pretreated liquid were prepared and their effect on Acremonium cellulase activity was monitored using Avicel. Xylo-oligomers and tannic acid were identified as powerful inhibitors of Acremonium cellulase, and the lowest hydrolysis rate of Avicel of 0.18g/g-glucose released/L/h was obtained from tannic acid.
Combined pretreatment with hot compressed water and wet disk milling opened up oil palm biomass structure resulting in enhanced enzymatic digestibility

Zakaria MR, Hirata S, Fujimoto S, Hassan MA

Bioresour Technol, 2015 Oct;193:128-34.
PMID: 26125612 DOI: 10.1016/j.biortech.2015.06.074

Combined pretreatment with hot compressed water and wet disk milling was performed with the aim to reduce the natural recalcitrance of oil palm biomass by opening its structure and provide maximal access to cellulase attack. Oil palm empty fruit bunch and oil palm frond fiber were first hydrothermally pretreated at 150-190° C and 10-240 min. Further treatment with wet disk milling resulted in nanofibrillation of fiber which caused the loosening of the tight biomass structure, thus increasing the subsequent enzymatic conversion of cellulose to glucose. The effectiveness of the combined pretreatments was evaluated by chemical composition changes, power consumption, morphological alterations by SEM and the enzymatic digestibility of treated samples. At optimal pretreatment process, approximately 88.5% and 100.0% of total sugar yields were obtained from oil palm empty fruit bunch and oil palm frond fiber samples, which only consumed about 15.1 and 23.5 MJ/kg of biomass, respectively.
Fulltext Optimal placement of unified power flow controllers to improve dynamic voltage stability using power system variable based voltage stability indices

Albatsh FM, Ahmad S, Mekhilef S, Mokhlis H, Hassan MA

PLoS One, 2015;10(4):e0123802.
PMID: 25874560 DOI: 10.1371/journal.pone.0123802

This study examines a new approach to selecting the locations of unified power flow controllers (UPFCs) in power system networks based on a dynamic analysis of voltage stability. Power system voltage stability indices (VSIs) including the line stability index (LQP), the voltage collapse proximity indicator (VCPI), and the line stability index (Lmn) are employed to identify the most suitable locations in the system for UPFCs. In this study, the locations of the UPFCs are identified by dynamically varying the loads across all of the load buses to represent actual power system conditions. Simulations were conducted in a power system computer-aided design (PSCAD) software using the IEEE 14-bus and 39- bus benchmark power system models. The simulation results demonstrate the effectiveness of the proposed method. When the UPFCs are placed in the locations obtained with the new approach, the voltage stability improves. A comparison of the steady-state VSIs resulting from the UPFCs placed in the locations obtained with the new approach and with particle swarm optimization (PSO) and differential evolution (DE), which are static methods, is presented. In all cases, the UPFC locations given by the proposed approach result in better voltage stability than those obtained with the other approaches.
Production of organic acids from kitchen wastes

Loh CW, Fakhru'l-Razi A, Hassan MA, Karim MI

Artif Cells Blood Substit Immobil Biotechnol, 1999 Sep-Nov;27(5-6):455-9.
PMID: 10595448

This study involves the production of short-chain organic acids from kitchen wastes as intermediates for the production of biodegradable plastics. Flasks, without mixing were used for the anaerobic conversion of the organic fraction of kitchen wastes into short-chain organic acids. The influence of pH, temperature and addition of sludge cake on the rate of organic acids production and yield were evaluated. Fermentations were carried out in an incubator at different temperatures controlled at 30 degrees C. 40 degrees C, 50 degrees C, 60 degrees C and uncontrolled at room temperature. The pH was also varied at pH 5, 6, 7, and uncontrolled pH. 1.0 M phosphate buffer was used for pH control, and 1.0 M HCl and 1.0 M NaOH were added when necessary. Sludge cake addition enhanced the rate of maximum acids production from 4 days to 1 day. The organic acids produced were maximum at pH 7 and 50 degrees C i.e., 39.84 g/l on the fourth day of fermentation with a yield of 0.87 g/g soluble COD consumed, and 0.84 g/g TVS. The main organic acid produced was lactic acid (65-85%), with small amounts of acetic (10-30%), propionic (5-10%), and butyric (5-20%) acids. The results of this study showed that kitchen wastes could be fermented to high concentration of organic acids, which could be used as substrates for the production of biodegradable plastics.
Degumming of crude palm oil by membrane filtration

Ong KK, Fakhru'l-Razi A, Baharin BS, Hassan MA

Artif Cells Blood Substit Immobil Biotechnol, 1999 Sep-Nov;27(5-6):381-5.
PMID: 10595436

The application of membrane separation in palm oil refining process has potential for energy and cost savings. The conventional refining of crude palm oil results in loss of oil and a contaminated effluent. Degumming of crude palm oil by membrane technology is conducted in this study. The objective of this research is to study the feasibility of membrane filtration for the removal of phospholipids in the degumming of crude palm oil, including analyses of phosphorus content, carotene content free fatty acids (as palmitic acid), colour and volatile matter. A PCI membrane module was used which was equipped with polyethersulfone membranes having a molecular weight cut off of 9,000 (type ES209). In this study, phosphorus content was the most important parameter monitored. The membrane effectively removed phospholipids resulting in a permeate with a phosphorus content of less than 0.3 ppm The percentage removal of phosphorus was 96.4% and was considered as a good removal. Lovibond colour was reduced from 27R 50Y to 20R 30Y. The percentage removal of carotene was 15.8%. The removal of colour was considered good but the removal of carotene was considered insignificant by the membrane. Free fatty acids and volatile matter were not removed. Typical of membrane operations, the permeate flux decreased with time and must be improved in order to be adopted on an industrial scale. Membrane technology was found to have good potential in crude palm oil degumming. However, an appropriate method has to be developed to clean the membranes for reuse.
Feasibility study on the utilization of rubber latex effluent for producing bacterial biopolymers

Tang SN, Fakhru'l-Razi A, Hassan MA, Karim MI

Artif Cells Blood Substit Immobil Biotechnol, 1999 Sep-Nov;27(5-6):411-6.
PMID: 10595441

Rubber latex effluent is a polluting source that has a high biochemical oxygen demand (BOD). It is estimated that about 100 million liters of effluent are discharged daily from rubber processing factories. Utilization of this effluent such as the use of a coupled system not only can reduce the cost of treatment but also yield a fermentation feedstock for the production of bioplastic. This study initially was carried out to increase the production of organic acids by anaerobic treatment of rubber latex effluent. It was found that through anaerobic treatment the concentration of organic acids did not increase. Consequently, separation of organic acids from rubber latex effluent by anion exchange resin was examined as a preliminary study of recovering acetic and propionic acids. However, the suspended solids (SS) content in the raw effluent was rather high which partially blocked the ion-exchange columns. Lime was used to remove the SS in the rubber latex effluent. After the lime precipitation process, organic acids were found to adsorb strongly onto the anion exchange resin. Less adsorption of organic acids onto the resin was observed before the lime precipitation. This was probably due to more sites being occupied by colloidal particles on the resin thus inhibiting the adsorption of organic acids. The initial concentration of organic acids in the raw effluent was 3.9 g/L. After ion exchange, the concentration of the organic acids increased to 27 g/L, which could be utilized for production of polyhydroxyalkanoates (PHA). For PHA accumulation stage, concentrated rubber latex effluent obtained from ion exchange resins and synthetic acetic acid were used as the carbon source. Quantitative analyses from fed batch culture via HPLC showed that the accumulation of PHA in Alcaligenes eutrophus was maximum with a concentration of 1.182 g/L when cultivated on synthetic acetic acid, corresponding to a yield of 87% based on its cell dry weight. The dry cell weight increased from 0.71 to 1.67 g/L. On the other hand, using concentrated rubber latex effluent containing acetic and propionic acids resulted in reduced PHA content by dry weight (14%) but the dry cell weight increased from 0.49 to 1.30 g/L. The results clearly indicated that the cells grow well in rubber latex effluent but no PHA was accumulated. This could be due to the high concentration of propionic acid in culture broth or other factors such as heavy metals. Thus further work is required before rubber latex effluent can be utilized as a substrate for PHA production industrially.
The application of a computerised recording system to smallholder dairy extension in Malaysia

Abu Hassan MA, Kamaruddin MI, Pharo HJ

Acta Vet Scand Suppl, 1988;84:110-2.
PMID: 3232593
Prophylactic Onlay Mesh Repair (POMR) Versus Primary Suture Repair (PSR) for Prevention of Incisional Hernia (IH) After Abdominal Wall Surgery: A Systematic Review and Meta-analysis

Hassan MA, Yunus RM, Khan S, Memon MA

World J Surg, 2021 10;45(10):3080-3091.
PMID: 34279690 DOI: 10.1007/s00268-021-06238-6

BACKGROUND: With many different operative techniques in use to reduce the incidence of incisional hernias (IH) following a midline laparotomy, there is no consensus among the clinicians on the efficacy and safety of any particular repair technique. This meta-analysis compares the prophylactic onlay mesh repair (POMR) and primary suture repair (PSR) for the incidence of IH.
METHODS: A meta-analysis and systematic review of MEDLINE, PubMed Central (via PubMed), Embase (via Ovid), SCOPUS, ScienceDirect, Google Scholar, SCI and Cochrane Library databases were undertaken. Seven randomized controlled trials assessing the outcomes of PSR and POMR were analyzed in accordance with the PRISMA statement. The risk of bias was assessed using the Rob2 tool.
RESULTS: According to the pooled analysis, POMR significantly reduced the incidence of IH compared to the PSR (OR 5.82 [95% CI 2.69, 12.58] P
Fulltext Towards health monitoring using remote heart rate measurement using digital camera: A feasibility study

Hassan MA, Malik AS, Fofi D, Karasfi B, Meriaudeau F

Measurement : journal of the International Measurement Confederation, 2020 Jan;149:106804.
PMID: 32287815 DOI: 10.1016/j.measurement.2019.07.032

The paper presents a feasibility study for heart rate measurement using a digital camera to perform health monitoring. The feasibility study investigates the reliability of the state of the art heart rate measuring methods in realistic situations. Therefore, an experiment was designed and carried out on 45 subjects to investigate the effects caused by illumination, motion, skin tone, and distance variance. The experiment was conducted for two main scenarios; human-computer interaction scenario and health monitoring scenario. The human-computer scenario investigated the effects caused by illumination variance, motion variance, and skin tone variance. The health monitoring scenario investigates the feasibility of health monitoring at public spaces (i.e. airports, subways, malls). Five state of the art heart rate measuring methods were re-implemented and tested with the feasibility study database. The results were compared with ground truth to estimate the heart rate measurement error. The heart rate measurement error was analyzed using mean error, standard deviation; root means square error and Pearson correlation coefficient. The findings of this experiment inferred promising results for health monitoring of subjects standing at a distance of 500 cm.
Promotion of a green economy with the palm oil industry for biodiversity conservation: A touchstone toward a sustainable bioindustry

Sakai K, Hassan MA, Vairappan CS, Shirai Y

J Biosci Bioeng, 2022 Feb 09.
PMID: 35151536 DOI: 10.1016/j.jbiosc.2022.01.001

Palm oil is a representative and important biomass, not only as the most edible vegetable oil consumed worldwide, but also as a material for chemicals and biofuels. Despite the potential sustainability of the palm oil industry, it has conventionally emitted excess greenhouse gases, waste materials, and wastewater, brought land use change, thus affecting the natural environment. Therefore, the successful development of a sustainable palm oil industry is a touchstone for promoting the bioeconomy. Here, we first review the concept of the bioeconomy and the positive and negative aspects of the palm oil industry. Then, we consider solutions for introducing a green economy into the palm oil industry, such that it may coexist with biodiversity and environmental conservation toward the Sustainable Development Goals.
Fulltext SWCNT Network-FET Device for Human Serum Albumin Detection

Yahya I, Hassan MA, Maidin NNM, Mohamed MA

Sensors (Basel), 2022 Oct 26;22(21).
PMID: 36365910 DOI: 10.3390/s22218212

A thin film of single-walled carbon nanotube (SWCNT) network field-effect transistor (FET) was fabricated by a simple, fast, and reliable deposition method for electronic applications. This study aims to develop a method for fabricating a thin film of random SWCNTs to be used as a transducer to detect human serum albumin (HSA) in biosensor applications. The random SWCNT network was deposited using the airbrush technique. The morphology of the CNT network was examined by utilising atomic force microscopy (AFM) and field-emission scanning electron microscopy (FESEM), while electrical characteristics were analysed using three-terminal IV measurements. The thin film (SWCNT network) was applied as a transducer to detect human serum albumin (HSA) based on its covalent interaction with antibodies. HSA plays a significant part in the physiological functions of the human body. The surface alteration of the SWCNTs was verified using Fourier transform infrared (FTIR) spectroscopy. Electrical current-voltage measurements validated the surface binding and HSA detection. The biosensor linearly recorded a 0.47 fg/mL limit of detection (LOD) and a high sensitivity of 3.44 μA (g/mL)-1 between 1 fg/mL and 10 pg/mL. This device can also be used to identify a genuine HSA despite interference from other biomolecules (i.e., bovine serum albumin (BSA)), thus demonstrating the random SWCNT-FET immunosensor ability to quantify HSA in a complex biological environment.
Novel health monitoring method using an RGB camera

Hassan MA, Malik AS, Fofi D, Saad N, Meriaudeau F

Biomed Opt Express, 2017 Nov 01;8(11):4838-4854.
PMID: 29188085 DOI: 10.1364/BOE.8.004838

In this paper we present a novel health monitoring method by estimating the heart rate and respiratory rate using an RGB camera. The heart rate and the respiratory rate are estimated from the photoplethysmography (PPG) and the respiratory motion. The method mainly operates by using the green spectrum of the RGB camera to generate a multivariate PPG signal to perform multivariate de-noising on the video signal to extract the resultant PPG signal. A periodicity based voting scheme (PVS) was used to measure the heart rate and respiratory rate from the estimated PPG signal. We evaluated our proposed method with a state of the art heart rate measuring method for two scenarios using the MAHNOB-HCI database and a self collected naturalistic environment database. The methods were furthermore evaluated for various scenarios at naturalistic environments such as a motion variance session and a skin tone variance session. Our proposed method operated robustly during the experiments and outperformed the state of the art heart rate measuring methods by compensating the effects of the naturalistic environment.
Kojic acid production by Aspergillus flavus using gelatinized and hydrolyzed sago starch as carbon sources

Rosfarizan M, Ariff AB, Hassan MA, Karim MI

Folia Microbiol (Praha), 1998;43(5):459-64.
PMID: 9867479

Direct conversion of gelatinized sago starch into kojic acid by Aspergillus flavus strain having amylolytic enzymes was carried out at two different scales of submerged batch fermentation in a 250-mL shake flask and in a 50-L stirred-tank fermentor. For comparison, fermentations were also carried out using glucose and glucose hydrolyzate from enzymic hydrolysis of sago starch as carbon sources. During kojic acid fermentation of starch, starch was first hydrolyzed to glucose by the action of alpha-amylase and glucoamylase during active growth phase. The glucose remaining during the production phase (non-growing phase) was then converted to kojic acid. Kojic acid production (23.5 g/L) using 100 g/L sago starch in a shake flask was comparable to fermentation of glucose (31.5 g/L) and glucose hydrolyzate (27.9 g/L) but in the 50-L fermentor was greatly reduced due to non-optimal aeration conditions. Kojic acid production using glucose was higher in the 50-L fermentor than in the shake flask.
Fulltext Recovery of glucose from residual starch of sago hampas for bioethanol production

Awg-Adeni DS, Bujang KB, Hassan MA, Abd-Aziz S

Biomed Res Int, 2013;2013:935852.
PMID: 23509813 DOI: 10.1155/2013/935852

Lower concentration of glucose was often obtained from enzymatic hydrolysis process of agricultural residue due to complexity of the biomass structure and properties. High substrate load feed into the hydrolysis system might solve this problem but has several other drawbacks such as low rate of reaction. In the present study, we have attempted to enhance glucose recovery from agricultural waste, namely, "sago hampas," through three cycles of enzymatic hydrolysis process. The substrate load at 7% (w/v) was seen to be suitable for the hydrolysis process with respect to the gelatinization reaction as well as sufficient mixture of the suspension for saccharification process. However, this study was focused on hydrolyzing starch of sago hampas, and thus to enhance concentration of glucose from 7% substrate load would be impossible. Thus, an alternative method termed as cycles I, II, and III which involved reusing the hydrolysate for subsequent enzymatic hydrolysis process was introduced. Greater improvement of glucose concentration (138.45 g/L) and better conversion yield (52.72%) were achieved with the completion of three cycles of hydrolysis. In comparison, cycle I and cycle II had glucose concentration of 27.79 g/L and 73.00 g/L, respectively. The glucose obtained was subsequently tested as substrate for bioethanol production using commercial baker's yeast. The fermentation process produced 40.30 g/L of ethanol after 16 h, which was equivalent to 93.29% of theoretical yield based on total glucose existing in fermentation media.
Hydrothermal pretreatment enhanced enzymatic hydrolysis and glucose production from oil palm biomass

Zakaria MR, Hirata S, Hassan MA

Bioresour Technol, 2015 Jan;176:142-8.
PMID: 25460995 DOI: 10.1016/j.biortech.2014.11.027

The present works investigate hydrothermal pretreatment of oil palm empty fruit bunch and oil palm frond fiber in a batch tube reactor system with temperature and time range from 170 to 250°C and 10 to 20min, respectively. The behavior of soluble sugars, acids, furans, and phenols dramatically changed over treatment severities as determined by HPLC. The cellulose-rich treated solids were analyzed by SEM, WAXD, and BET surface area. Enzymatic hydrolysis was performed from both pretreated slurries and washed solid, and data obtained suggested that tannic acid derived from lignin degradation was a potential cellulase inhibitor. Both partial removal of hemicellulose and migration of lignin during hydrothermal pretreatment caused structural changes on the cellulose-hemicellulose-lignin matrix, resulting in the opening and expansion of specific surface area and pore volume. The current results provided important factors that maximize conversion of cellulose to glucose from oil palm biomass by hydrothermal process.
Combined pretreatment using alkaline hydrothermal and ball milling to enhance enzymatic hydrolysis of oil palm mesocarp fiber

Zakaria MR, Hirata S, Hassan MA

Bioresour Technol, 2014 Oct;169:236-43.
PMID: 25058299 DOI: 10.1016/j.biortech.2014.06.095

Hydrothermal pretreatment of oil palm mesocarp fiber was conducted in tube reactor at treatment severity ranges of log Ro = 3.66-4.83 and partial removal of hemicellulose with migration of lignin was obtained. Concerning maximal recovery of glucose and xylose, 1.5% NaOH was impregnated in the system and subsequent ball milling treatment was employed to improve the conversion yield. The effects of combined hydrothermal and ball milling pretreatments were evaluated by chemical composition changes by using FT-IR, WAXD and morphological alterations by SEM. The successful of pretreatments were assessed by the degree of enzymatic digestibility of treated samples. The highest xylose and glucose yields obtained were 63.2% and 97.3% respectively at cellulase loadings of 10 FPU/g-substrate which is the highest conversion from OPMF ever reported.

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