The aim of this study is to develop bioplastic film from a combination of two biopolymers of same
source, namely banana peel and corn starch. Five banana peel films (BP film) were prepared with
different concentrations of corn starch (1% up to 5%) as co-biopolymer and film without corn
starch acted as a control. The films were carried out with several durability tests and
characterization analyses. Based on the results obtained, the BP film with 4% corn starch gave the
highest tensile strength 34.72 N/m2 compared to other samples. The water absorption test showed
that BP films with 3% corn starch were resistant to water uptake by absorbing water up to 60.65%.
In terms of characterization, spectra of Fourier Transform Infrared Spectroscopy (FTIR) obtained
for BP control film and BP film with 4% corn starch were comparable with most of the peaks were
present. The thermal analysis by differential screening calorimetric (DSC) detected the melting
temperature for both BP control film and BP film with 4% corn respectively at Tonset of 54.41°C
and 67.83°C. Overall, combination of starches from two different sources can be used as an
alternative in producing bioplastics.
The growth of microorganism on substrates, whether toxic or not usually exhibits sigmoidal
pattern. This sigmoidal growth pattern can be modelled using primary models such as Logistic,
modified Gompertz, Richards, Schnute, Baranyi-Roberts, Von Bertalanffy, Buchanan threephase
and Huang. Previously, the modified Gompertz model was chosen to model the growth of
Burkholderia sp. strain Neni-11 on acrylamide, which shows a sigmoidal curve. The modified
Gompertz model relies on the ordinary least squares method, which in turn relies heavily on
several important assumptions, which include that the data does not show autocorrelation. In this
work we perform statistical diagnosis test to test for the presence of autocorrelation using the
Durbin-Watson test and found that the model was adequate and robust as no autocorrelation of
the data was found.
Petroleum hydrocarbons remain as the major contaminants that could be found across the world.
Remediation approach through the utilisation of microbes as the bioremediation means widely
recognised due to their outstanding values. As a result, scientific reports on the isolation and
identification of new hydrocarbon-degrading strains were on the rise. Colourimetric-based assays
are one of the fastest methods to identify the capability of hydrocarbon-degrading strains in both
qualitative and quantitative assessment. In this study, the hydrocarbon-degrading potential of
nine bacterial isolates was observed via 2,6-dichlorophenolindophenol (DCPIP) test. Two potent
diesel-utilising isolates show a distinctive tendency to utilise aromatic (ADL15) and aliphatic
(ADL36) hydrocarbons. Both isolates prove to be a good candidate for bioremediation of wide
range of petroleum hydrocarbon components.
Bioremediation is a new green economic approach in providing solutions for cleaning up
contaminated sites. Phytoremediation uses plants as a tool for remediation purposes. The usage
of plant species offers higher potential solution to remediate heavy metal contaminated sites.
This study aimed on screening potential plant species for phytoremediation of heavy metal
contaminated water. The potential of three aquatic macrophytes species (Eichorrnia crassipes,
Pistia stratiotes and Ipomoea aquatica) for chromium and nickel phytoremediations was tested.
The plants were exposed for 10 days under hydroponic conditions in heavy metal contaminated
water. E. crassipes showed the highest chromium and nickel concentrations in its biomass, 1.60
and 2.40 μg/L respectively. Meanwhile, P. stratiotes had chromium and nickel concentrations
detected at 0.89 and 0.081 μg/L, respectively; chromium and nickel concentrations of I.
aquatica detected were, 0.49 and 0.08 μg/L, respectively. The ability of these plants to
accumulate heavy metals and survived throughout the experiment demonstrates the potential of
these plants to remediate metal-enriched water. Among the three tested aquatic plants, E.
crassipes was proven to be the most suitable plant species that can phytoremediate heavy metal
contaminated water followed by P. stratiotes and I. aquatica.
The trend of global under-five child mortality rate showed a steady decrease at -3.14% per year
approaching the United Nations millennium development goals target (-4.0% reduction per year
from 1990-2015). This rate is usually inversely proportional to the nations GDP. However, the
progress in the recent years (2006-2012) in Malaysia has raised a disturbing concern with an
annual average increment of +0.81% per year. Thus, there is an urgent need to identify the
reasons of such risky ecosystem. By analyzing recorded data on river pollution from 2007-
2012, we found that river pollution has high correlation with the gross domestic products, not at
the same year, but for two years prior that indicates a lag time. We also found that under-five
child mortality has strong correlation with the river pollutions. The development-pollutionhealth
triangular cycle needs to be put in a balanced to ensure the nation prosperity and
sustainability of the nation.
Mass production of fish broodstock with high quality eggs requires the knowledge on the chemical composition and physiochemical properties of vitellogenin (Vtg) during ovulation. Vtg is an egg yolk precursor phospholipoglycoprotein, and has been analysed to evaluate the reproductive conditions and determine the spawning period in captive and wild fish. In this study, Vtg was induced in male H. nemurus through three intramuscular injections of 17-estradiol (E2). The Vtg was purified from the serum using gel filtration chromatography and the purified protein was reduced via SDS-PAGE. One major polypeptide corresponding to 130 kDa was observed. Vtg identification was done using peptide mass fingerprint (PMF) from the trypsin digestion of male H. nemurus Vtg induced with E2. The sequence homology of H. nemurus AYLAGAAADVLEVGVR matched the Vtg of other fish species when analysed using MALDI-TOF. Vtg was confirmed by MASCOT at 95% significant level. The potential protein that controls the reproductive process and oocyte development isolated from this study was discussed to understand the structure and function of Vtg.
Biosurfactants are surface active compounds and amphiphatic in nature which consist of
hydrophilic head and hydrophobic tail accumulating at the interphase of two immiscible liquid
with different polarity. A study was conducted to investigate the effectiveness of sunflower oil in
the production of rhamnolipids (RLs) by locally isolated Pseudomonas aeruginosa in shake flask
fermentation. In this process, four different fermentation treatments were done for seven days at
30°C and 180 rpm. Sampling was carried out in time intervals of 24 h followed by monitoring of
cell growth and biosurfactants production. Colorimetric Orcinol analysis was used for
determination of RLs concentrations (g/L). The RLs were studied for emulsification activity
using emulsification index (E24%) methods. In addition, oil displacement activity and thermal
stability were also studied (4-120°C). All treatments allow the growth of P. aeruginosa and the
utilization of sunflower oil as carbon source and glucose as growth initiator were observed to be
the best strategy for maximum RLs production. The maximum RLs production was achieved
after 120 h with 3.18 g/L of RLs. Diesel shows the highest emulsification activity among the
substrate tested ranging from 55.56% - 60.00%. The oil displacement activity was corresponding
to RLs concentration with stability up to 120°C (for 60 min). Therefore, from this research a
good potential of RLs that may provide good application for industry were produced.
Heavy metals pollution has become a great threat to the world. Since instrumental methods are
expensive and need skilled technician, a simple and fast method is needed to determine the
presence of heavy metals in the environment. In this work, a preliminary study was carried out
on the applicability of various local plants as a source of protease for the future development of
the inhibitive enzyme assay for heavy-metals. The crude proteases preparation was assayed using
casein as a substrate in conjunction with the Coomassie dye-binding assay. The crude protease
from the kesinai plant was found to be the most potent plant protease. The crude enzyme
exhibited broad temperature and pH ranges for activity and will be developed in the future as a
potential inhibitive assay for heavy metals.
Protein function depends greatly on its structure. Based on this principle, it is vital to study the
protein structure in order to understand its function. This study attempts to build the predicted
model of lipase gene in Rhodococcus sp. NAM81 using homology modelling method. The
predicted structure was then used to investigate the function of protein through several
bioinformatic tools. The DNA sequence of lipase gene was obtained from the Rhodococcus sp.
NAM81 genome scaffold. Blastx analysis showed 100% identity to the target enzyme andthe
appropriate template for homology modelling was determined using Blastp analysis. The 3D
protein structure was built using two homology modelling software, EsyPred3D and Swiss
Model Server. Both structures built obtained LGScore of greater than 4, which means they are
extremely good models according to ProQ validation criteria. Both structures also satisfied the
Ramachandran plot structure validation analysis. The predicted structures were 100% matched
with each other when superimposed with DaliLite pairwise. This shows that both structure
validation servers agreed on the same model. Structure analysis using ProFunc had found seven
motifs and active sites that indicate similar function of this protein with other known proteins.
Thus, this study has successfully produced a good 3D protein structure for the target enzyme.
Environmental pollution is one of the major concerns in the 21st century; where billions of tonnes
of harmful chemicals are produced by industries such as petroleum, paints, food, rubber, and
plastic. Phenol and its derivatives infiltrate the ecosystems and have become one of the top major
pollutants worldwide. This review covers the major aspects of immobilization of phenoldegrading
bacteria as a method to improve phenol bioremediation. The use of various forms of
immobilization matrices is discussed along with the advantages and disadvantages of each of the
immobilization matrices especially when environmental usage is warranted. To be used as a
bioremediation tool, the immobilized system must not only be effective, but the matrices must be
non-toxic, non-polluting and if possible non-biodegradable. The mechanical, biological and
chemical stability of the system is paramount for long-term activity as well as price is an
important factor when the very large scale is a concern. The system must also be able to tolerate
high concentration of other toxicants especially heavy metals that form as co-contaminants, and
most immobilized systems are geared towards this last aspect as immobilization provides
protection from other contaminants.
Phenolic compounds or phenols are a group of aromatic compounds that comprises a hydroxyl
group (OH) that is directly bonded to an aromatic ring. Phenols are injurious to organisms even
at even low concentrations with many of them are categorized as dangerous pollutants because of
their likely harm to human well-being. This review attempts to discuss the various merits and
demerits of immobilization matrices employed for phenol-degrading microorganisms’
immobilization. One of several key points of cellular immobilization is the capacity to protect
bioremediation agents towards toxic levels of specific toxicants and safeguarding from predatory
microorganisms. However, this shielding course of action should never impede the diffusion of
substrates into the pores of the immobilization structure. In the end the choice of a particular
immobilization method will strongly hinge on aspects of economy, safety and efficacy.
Molybdenum is an emerging pollutant. Bioremediation of this heavy metal is possible by the
mediation of Mo-reducing bacteria. These bacteria contain the Mo-reducing enzymes that can
conver toxic soluble molybdenum into molybdenum blue; a less soluble and less toxic form of the
metal. To date only the enzyme has been purified from only one bacterium. The aim of this study is
to purify the Mo-reducing enzyme from a previously isolated Mo-reducing bacterium Bacillus
pumilus strain Lbna using ammonium sulphate fractionation followed by ion exchange and then
gel filtration. Two clear bands were obtained after the gel filtration step with molecular weights
of 70 and 100 kDa. This indicates that further additional purification methods need to be used
to get a purified fraction. Hence, additional steps of chromatography such as hydroxyapatite or
chromatofocusing techniques can be applied in the future.
In this study, a novel glyphosate-degrading shows the ability to reduce molybdenum to
molybdenum blue. The enzyme from this bacterium was partially purified and partially
characterized to ascertain whether the Mo-reducing enzyme from this bacterium shows better or
lower efficiency in reducing molybdenum compared to other Mo-reducing bacterium that only
exhibits a single biotransformation activity. The enzyme was partially purified using ammonium
sulphate fractionation. The Vmax for the electron donating substrate or NADH was at 1.905 nmole
Mo blue/min while the Km was 6.146 mM. The regression coefficient was 0.98. Comparative
assessment with the previously characterized Mo-reducing enzyme from various bacteria showed
that the Mo-reducing enzyme from Burkholderia vietnamiensis strain AQ5-12 showed a lower
enzyme activity.
Investigation on in vivo effects of copper (Cu) on the ultrastructure of P. javanicus liver was
carried out using transmission electron microscopy (TEM). The addition of sublethal
concentration of 5 mg/L of Cu caused abnormalities on the bile canaliculi (BC) including
dilation and elongation compared to control and at lower concentrations of copper with a normal
round shape form. Findings from this study support an alternative histological assessment of the
effects of Cu concentration on P. javanicus liver.
In this work, a temporal monitoring work for heavy metals from an effluent discharge point in
the Juru Industrial Estate was carried out using the protease extracted from garlic (Allium
sativum) as the principal bioassay system. casein-Coomassie-dye binding assay method has
utilized this purpose. The periodic sampling results for one day of a location in the Juru
Industrial Estate showed temporal variation of copper concentration coinciding with garlic
protease inhibition with the highest concentrations of copper occurring between 12.00 and 16.00
hours of between 3 and 3.5 mg/L copper. The crude proteases extracted from Allium sativum
successfully detect temporal variation of copper form this location. In conclusion, this assay
method has the potential to be a rapid, sensitive, and economic inhibitive assay for the largescale
biomonitoring works for the heavy metal copper from this area.
Quinolines compounds are toxic pollutants. Their biodegradation by microbes represents a tool
for bioremediation. The growth of Klebsiella penumoniae on 2-methylquinoline shows typical
sigmoidal bacterial growth curves. Since there exists a variety of models for describing the
growth profile of microorganism such as logistic, Gompertz, Richards, Schnute, Baranyi-
Roberts, Von Bertalanffy, Buchanan three-phase and more recently Huang models, the growth
curves exhibit under such conditions would be an excellent study for finding the best model.
The Huang model was chosen as the best model based on statistical tests such as root-meansquare
error (RMSE), adjusted coefficient of determination (R2), bias factor (BF), accuracy
factor (AF) and corrected AICc (Akaike Information Criterion). Novel constants obtained from
the modelling exercise would be used for further secondary modelling.
Pollution in the environment is deteriorating the ecology due to human activities in a large array
of industrial and agricultural sectors. Bioassay of polluted waters using bioluminescent bacterium
has been touted as one of the most economical, rapid and sensitive tests. The growth of the
bacterium on seawater medium exhibited a typical sigmoidal profile. To extract important growth
parameters useful for further modelling exercise, various primary growth models were utilized in
this study such as Modified Logistic, modified Gompertz, modified Richards, modified Schnute,
Baranyi-Roberts, von Bertalanffy, Huang and the Buchanan three-phase model. The best
performance was Huang model with the lowest value for RMSE, AICc and the highest value for
adjusted R2. The AF and BF values were also excellent for the model with their values were the
closest to 1.0. The Huang parameters, which include A or Y0 (bacterial growth lower asymptote),
μm (maximum specific bacterial growth rate), l (lag time) and Ymax (bacterial growth upper
asymptote) were 7.866 (95% confidence interval of 7.850 to 7.883), 0.329 (95% confidence
interval of 0.299 to 0.359), 1.543 (95% confidence interval of 1.303 to 1.784) and 8.511 (95%
confidence interval of 0.299 to 0.359).
Molybdenum, an emerging pollutant, has being demonstrated recently to be toxic to
spermatogenesis in several animal model systems. Metal mines especially gold mine often use
cyanide and hence isolation of metal-reducing and cyanide-degrading bacteria can be useful for
the bioremediation of these pollutants. Preliminary screening shows that three cyanide-degrading
bacteria were able to reduce molybdenum to molybdenum blue (Mo-blue) when grown on a
molybdate low phosphate minimal salts media. Phylogenetic analyses of the 16S rRNA gene of
the best reducer indicates that it belongs to the Serratia genus. A variety of mathematical models
such as logistic, Gompertz, Richards, Schnute, Baranyi-Roberts, von Bertalanffy, Buchanan
three-phase and Huang were used to model molybdenum reduction, and the best model based on
statistical analysis was modified Gompertz with lowest values for RMSE and AICc, highest
adjusted R2 values, with Bias Factor and Accuracy Factor nearest to unity (1.0). The reduction
constants obtained from the model will be used to carry out secondary modelling to study the
effect of various parameters such as substrate, pH and temperature to molybdenum reduction.
The pollution of heavy metals and toxic xenobiotics has become a central issue worldwide.
Bioremediation of these toxicants are being constantly carried out using novel microbes.
Molybdenum reduction to molybdenum blue is a detoxification process and mathematical
modelling of the reduction process can reveal important parameters such as specific reduction
rate, theoretical maximum reduction and whether reduction at high molybdenum concentration
affected the lag period of reduction. The used of linearization method through the use of natural
logarithm transformation, although popular, is inaccurate and can only give an approximate
value for the sole parameter measured; the specific growth rate. In this work, a variety of
models for such as logistic, Gompertz, Richards, Schnute, Baranyi-Roberts, Von Bertalanffy,
Buchanan three-phase and more recently Huang were utilized for the first time to obtain values
for the above parameters or constants. The modified Gompertz model was the best model in
modelling the Mo-blue production curve from Serratia marcescens strain DR.Y10 based on
statistical tests such as root-mean-square error (RMSE), adjusted coefficient of determination
(R2), bias factor (BF), accuracy factor (AF) and corrected AICc (Akaike Information Criterion).
Parameters obtained from the fitting exercise were maximum Mo-blue production rate (μm), lag
time (l) and maximal Mo-blue production (Ymax) of X (h-1), Y (h) and Z (nmole Mo-blue),
respectively. The application of primary population growth models in modelling the Moblue
production rate from this bacterium has become a successful undertaking. The model
may also be used in other heavy metals detoxification processes. The parameters
constants extracted from this work will be a substantial help for the future development
of further secondary models.
Bacterial based remediation of environmental toxicants is a promising innovative technology
for molybdenum pollution. To date, the enzyme responsible for molybdate reduction to Moblue
from bacteria show that the Michaelis-Menten constants varies by one order of magnitude.
It is important that the constants from newer enzyme sources be characterized so that a
comparison can be made. The aim of this study is to characterize kinetically the enzyme from a
previously isolated Mo-reducing bacterium; Bacillus pumilus strain Lbna. The maximum
activity of this enzyme occurred at pH 5.5 and in between 25 and 35 oC. The Km and Vmax of
NADH were 6.646 mM and 0.057 unit/mg enzyme, while the Km and Vmax of LPPM were 3.399
mM and 0.106 unit/mg enzyme. The results showed that the enzyme activity for Bacillus
pumilus strain Lbna were inhibited by all heavy metals used. Zinc, copper, silver, chromium,
cadmium and mercury all caused more than 50% inhibition to the Mo-reducing enzyme activity
with copper being the most potent with an almost complete inhibition of enzyme activity
observed.