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Response to invited commentary: Vitamin D3 supplementation for 8 weeks leads to improved haematological status following the consumption of an iron-fortified breakfast cereal: a double-blind randomised controlled trial in iron-deficient women

Ahmad Fuzi SF, Mushtaq S

Br J Nutr, 2021 Mar 14;125(5):598-600.
PMID: 32406343 DOI: 10.1017/S0007114520001683
Simplified feeding strategies for the fed-batch cultivation of Kluyveromyces lactis GG799 for enhanced recombinant xylanase production

Fuzi SF, Razali F, Jahim JM, Rahman RA, Illias RM

Bioprocess Biosyst Eng, 2014 Sep;37(9):1887-98.
PMID: 24633311 DOI: 10.1007/s00449-014-1163-z

A xylanase gene (xyn2) from Trichoderma reesei ATCC 58350 was previously cloned and expressed in Kluyveromyces lactis GG799. The production of the recombinant xylanase was conducted in a developed medium with an optimised batch and with fed-batches that were processed with glucose. The glucose served as a carbon source for cell growth and as an inducer for xylanase production. In a 1-L batch system, a glucose concentration of 20 g L(-1) and 80 % dissolved oxygen were found to provide the best conditions for the tested ranges. A xylanase activity of 75.53 U mL(-1) was obtained. However, in the batch mode, glucose depletions reduced the synthesis of recombinant xylanase by K. lactis GG799. To maximise the production of xylanase, further optimisation was performed using exponential feeding. We investigated the effects of various nitrogen sources combined with the carbon to nitrogen (C/N) molar ratio on the production of xylanase. Of the various nitrogen sources, yeast extract was found to be the most useful for recombinant xylanase production. The highest xylanase production (110.13 U mL(-1)) was measured at a C/N ratio of 50.08. These conditions led to a 45.8 % increase in xylanase activity compared with the batch cultures. Interestingly, the further addition of 500 g L(-1) glucose led to a 6.2-fold increase (465.07 U mL(-1)) in recombinant xylanase activity. These findings, together with those of the exponential feeding strategy, indicate that the composition of the C/N molar ratio has a substantial impact on recombinant protein production in K. lactis.
Fulltext Structural adaptation of cold-active RTX lipase from Pseudomonas sp. strain AMS8 revealed via homology and molecular dynamics simulation approaches

Mohamad Ali MS, Mohd Fuzi SF, Ganasen M, Abdul Rahman RN, Basri M, Salleh AB

Biomed Res Int, 2013;2013:925373.
PMID: 23738333 DOI: 10.1155/2013/925373

The psychrophilic enzyme is an interesting subject to study due to its special ability to adapt to extreme temperatures, unlike typical enzymes. Utilizing computer-aided software, the predicted structure and function of the enzyme lipase AMS8 (LipAMS8) (isolated from the psychrophilic Pseudomonas sp., obtained from the Antarctic soil) are studied. The enzyme shows significant sequence similarities with lipases from Pseudomonas sp. MIS38 and Serratia marcescens. These similarities aid in the prediction of the 3D molecular structure of the enzyme. In this study, 12 ns MD simulation is performed at different temperatures for structural flexibility and stability analysis. The results show that the enzyme is most stable at 0°C and 5°C. In terms of stability and flexibility, the catalytic domain (N-terminus) maintained its stability more than the noncatalytic domain (C-terminus), but the non-catalytic domain showed higher flexibility than the catalytic domain. The analysis of the structure and function of LipAMS8 provides new insights into the structural adaptation of this protein at low temperatures. The information obtained could be a useful tool for low temperature industrial applications and molecular engineering purposes, in the near future.
Growth of microalgae Botryococcus sp. in domestic wastewater and application of statistical analysis for the optimization of flocculation using alum and chitosan

Gani P, Mohamed Sunar N, Matias-Peralta H, Abdul Latiff AA, Mohamad Fuzi SF

Prep Biochem Biotechnol, 2017 Apr 21;47(4):333-341.
PMID: 27737612 DOI: 10.1080/10826068.2016.1244686

Microalga biomass has been recognized as a sustainable bio-product to replace terrestrial biomass in biofuel production. The microalga industry has high operating costs, specifically on harvesting and biomass recovery. Therefore, the development of an efficient harvesting method is crucial to the minimization of production cost. A statistical analysis through response surface methodology was used to investigate the optimization of harvesting efficiency using alum and chitosan as a coagulant. Growth rate and biomass productivity were also determined. This research revealed that the harvesting efficiency using alum was 99.3%, with optimum dosage and pH of 177.74 mg L(-1) and 8.24, respectively. Chitosan achieved 94.2% biomass recovery at an optimal dosage of 169.95 mg L(-1) at pH of 12. Moreover, Botryococcus sp. achieved the maximum growth of 0.7551 µmax d(-1), with an average total biomass productivity of 9.81 mg L(-1) d(-1) in domestic wastewater. Overall, this study shows that both alum and chitosan coagulants have great potential for efficient microalgal biomass recovery. It suggests that domestic wastewater as a potential growth medium for the large-scale production of microalga biomass.
Fulltext Exploration of Food-Seeking Behaviour, Food Preparation, and Restrictions to Sufficient Food among the Jahai Sub-Tribe (Indigenous People) in Gerik, Malaysia

Gan WY, Sulaiman N, Law LS, Zalbahar N, Ahmad Fuzi SF, Wilkes MA

Int J Environ Res Public Health, 2020 01 04;17(1).
PMID: 31947907 DOI: 10.3390/ijerph17010348

Access to food has been determined to be a factor that strongly influences the dietary intake and eating habit of indigenous people (Orang Asli, OA). This study explored food acquisition and the barriers in obtaining traditional and market foods among the Jahai subtribe (within the Negrito ethnic group) via a qualitative approach. Twenty-eight OA (14 males and 14 females) from Gerik, Perak, Malaysia participated in four focus group discussions (FGDs; two male-only and two female-only groups; seven informants in each FGD). Thematic analysis was adopted to analyse the gathered data. The results found that the Jahai applied both modern (buying and receiving food assistance) and traditional (gathering, farming, fishing, and hunting) methods in obtaining food. The barriers they faced in gathering sufficient food supply included low purchasing power, high demand for food, high transport costs, depletion of food supplies in their surroundings, threats from wild animals on the farm and during food searching activities, and food sharing. Food preparation methods practiced by the OA include roasting and grilling, frying, simmering (gulai), and boiling. In conclusion, this study enhances our understanding of the dietary behaviour of the Jahai subtribe and highlights restricted food access among the OA, which demands urgent attention.
A 1-h time interval between a meal containing iron and consumption of tea attenuates the inhibitory effects on iron absorption: a controlled trial in a cohort of healthy UK women using a stable iron isotope

Ahmad Fuzi SF, Koller D, Bruggraber S, Pereira DI, Dainty JR, Mushtaq S

Am J Clin Nutr, 2017 Dec;106(6):1413-1421.
PMID: 29046302 DOI: 10.3945/ajcn.117.161364

Background: Tea has been shown to be a potent inhibitor of nonheme iron absorption, but it remains unclear whether the timing of tea consumption relative to a meal influences iron bioavailability.Objective: The aim of the study was to investigate the effect of a 1-h time interval of tea consumption on nonheme iron absorption in an iron-containing meal in a cohort of iron-replete, nonanemic female subjects with the use of a stable isotope (57Fe).Design: Twelve women (mean ± SD age: 24.8 ± 6.9 y) were administered a standardized porridge meal extrinsically labeled with 4 mg 57Fe as FeSO4 on 3 separate occasions, with a 14-d time interval between each test meal (TM). The TM was administered with water (TM-1), with tea administered simultaneously (TM-2), and with tea administered 1 h postmeal (TM-3). Fasted venous blood samples were collected for iron isotopic analysis and measurement of iron status biomarkers. Fractional iron absorption was estimated by the erythrocyte iron incorporation method.Results: Iron absorption was 5.7% ± 8.5% (TM-1), 3.6% ± 4.2% (TM-2), and 5.7% ± 5.4% (TM-3). Mean fractional iron absorption was found to be significantly higher (2.2%) when tea was administered 1 h postmeal (TM-3) than when tea was administered simultaneously with the meal (TM-2) (P = 0.046). An ∼50% reduction in the inhibitory effect of tea (relative to water) was observed, from 37.2% (TM-2) to 18.1% (TM-3).Conclusions: This study shows that tea consumed simultaneously with an iron-containing porridge meal leads to decreased nonheme iron absorption and that a 1-h time interval between a meal and tea consumption attenuates the inhibitory effect, resulting in increased nonheme iron absorption. These findings are not only important in relation to the management of iron deficiency but should also inform dietary advice, especially that given to those at risk of deficiency. This trial was registered at clinicaltrials.gov as NCT02365103.
An 8-Week Vitamin D3-Fortified Fruit Drink Supplementation Increases Serum Ferritin Concentration: A Randomized Controlled Trial in Malaysian Women With Low Iron Stores

Suhaimi NA, Loh SP, Ab Manan N, Zalbahar N, Mohamad Alwi MN, Ahmad Fuzi SF

J Acad Nutr Diet, 2024 Feb 24.
PMID: 38408566 DOI: 10.1016/j.jand.2024.02.013

BACKGROUND: There is limited randomized controlled trial evidence to support the association between vitamin D deficiency and anemia risk, highlighting the necessity for further investigations into the role of vitamin D in influencing iron status.
OBJECTIVE: The aim of this study was to determine the effect of vitamin D3-fortified fruit drink consumption (4,000 IU) on vitamin D and iron status biomarkers among iron-deficient women (serum ferritin of <20 μg/L [to convert μg/L ferritin to ng/mL, multiply by 1]).
DESIGN: An 8-week double-blind randomized controlled trial was conducted.
SUBJECTS/SETTING: A total of 45 healthy, nonpregnant, nonlactating subjects aged 18 through 40 years (mean [SD] 25.3 [4.6] years) were included in the study, excluding those who donated blood 6 months prior, regularly consumed nutritional supplements, or had gastrointestinal or iron metabolic disorders.
INTERVENTION: Subjects were randomly assigned to receive either vitamin D3-fortified fruit drink or a placebo.
MAIN OUTCOME MEASURES: Measurements of 25-hydroxyvitamin D (25[OH]D), serum ferritin, high-sensitivity C-reactive protein, and full blood count concentrations were obtained at baseline, interim, and post intervention.
STATISTICAL ANALYSES: A mixed model, repeated measures analysis of variance was used to analyze the intervention effect.
RESULTS: Attrition rate for the study was 13%, with 6 dropouts, and 39 subjects completed the study. Daily consumption of vitamin D3-fortified fruit drink in the intervention group resulted in significant increases in 25(OH)D and serum ferritin concentrations compared with the placebo group. The intervention group showed significantly higher mean (SD) changes (Δ) in both 25(OH)D (Δ 76.4 [30.2] nmol/L [to convert nmol/L 25(OH)D to ng/mL, multiply by .4] vs Δ -1.3 [10.7] nmol/L; P = .001) and serum ferritin concentrations (Δ 2.2 [4.2] μg/L vs Δ -0.3 [3.4] μg/L; P = .048) between baseline and post intervention. The other iron status biomarkers were not affected by the intervention.
CONCLUSIONS: Our study found that daily vitamin D3-fortified fruit drink supplementation for 8 weeks effectively improved 25(OH)D and iron stores, indicated by increased serum ferritin concentrations, in iron-deficient women. Further research is needed to evaluate its safety, efficacy, feasibility, and optimal food fortification in diverse populations.
Evaluation on microalgae for the production of bio-chemicals and electricity

Khairuddin F, Zaharah Mohd Fuzi SF, Ahmad A, Oon LK, Bokhari A, Dailin DJ, et al.

Chemosphere, 2024 Feb;350:141007.
PMID: 38141667 DOI: 10.1016/j.chemosphere.2023.141007

Recent advancement in biophotovoltaic systems using microalgae, coupled with biorefinery approach, would improve economy-feasibility in production. The major concern is its commercial strength in terms of scalability, strain selection and extraction procedure cost. It must compete with conventional feedstocks such as fossil fuels. This project proposes to enhance the economic feasibility of microalgae-based biorefinery by evaluating their performance for bio-electricity, bio-diesel and carotenoids production in a single cycle. The first part of the study was to construct and select a Bio-bottle Voltaic (BBV) device that would allow microalgae to grow and produce bioproducts, as well as generate the maximum current output reading derived from the microalgae's photosynthesis process. The second phase consisted of a 25-day investigation into the biorefinery performance of six different microalgal species in producing bio-electricity, bio-diesel and carotenoid in a prototype BBV device. The prototype BBV device with aluminium foil and pencil lead as its anode and cathode produced the highest carotenoid and biodiesel component production from the two microalgae tested, according to the results of the first phase of the experiment. In the second portion of the study, Scenedesmus dimorphus and Chlorella vulgaris were identified as the two microalgae most capable of maintaining their growth throughout the experiment. The maximum current reading observed for C. vulgaris was 653 mV. High Performance Liquid Chromatography analysis showed four major carotenoid compounds found which were Neoxanthin, Cantaxanthin, Astaxanthin and 9-cis antheraxanthin, and the highest carotenoid producer was C. vulgaris which recorded at 1.73 μg/mL. C. vulgaris recorded as the most alkanes producer with 22 compounds detected and Heptacosane and Heneicosane as the two major biodiesel compounds found in the extracts. Evaluation of C. vulgaris data showed that it has enormous potential for microalgal biorefinery candidates. Further ongoing research and development efforts for C. vulgaris will improve the economic viability of microalgae-based industries and reduce reliance on depleted fossil fuels.
A comparative genomic analysis of the alkalitolerant soil bacterium Bacillus lehensis G1

Noor YM, Samsulrizal NH, Jema'on NA, Low KO, Ramli AN, Alias NI, et al.

Gene, 2014 Jul 25;545(2):253-61.
PMID: 24811681 DOI: 10.1016/j.gene.2014.05.012

Bacillus lehensis G1 is a Gram-positive, moderately alkalitolerant bacterium isolated from soil samples. B. lehensis produces cyclodextrin glucanotransferase (CGTase), an enzyme that has enabled the extensive use of cyclodextrin in foodstuffs, chemicals, and pharmaceuticals. The genome sequence of B. lehensis G1 consists of a single circular 3.99 Mb chromosome containing 4017 protein-coding sequences (CDSs), of which 2818 (70.15%) have assigned biological roles, 936 (23.30%) have conserved domains with unknown functions, and 263 (6.55%) have no match with any protein database. Bacillus clausii KSM-K16 was established as the closest relative to B. lehensis G1 based on gene content similarity and 16S rRNA phylogenetic analysis. A total of 2820 proteins from B. lehensis G1 were found to have orthologues in B. clausii, including sodium-proton antiporters, transport proteins, and proteins involved in ATP synthesis. A comparative analysis of these proteins and those in B. clausii and other alkaliphilic Bacillus species was carried out to investigate their contributions towards the alkalitolerance of the microorganism. The similarities and differences in alkalitolerance-related genes among alkalitolerant/alkaliphilic Bacillus species highlight the complex mechanism of pH homeostasis. The B. lehensis G1 genome was also mined for proteins and enzymes with potential viability for industrial and commercial purposes.