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Fulltext Identification of local clinical Candida isolates using CHROMagar Candida™ as a primary identification method for various Candida species

Madhavan P, Jamal F, Chong PP, Ng KP

Trop Biomed, 2011 Aug;28(2):269-74.
PMID: 22041745

The objective of our study was to study the effectiveness of CHROMagar Candida™ as the primary identification method for various clinical Candida isolates, other than the three suggested species by the manufacturer. We studied 34 clinical isolates which were isolated from patients in a local teaching hospital and 7 ATCC strains. These strains were first cultured in Sabouraud dextrose broth (SDB) for 36 hours at 35ºC, then on CHROMagar plates at 30ºC, 35ºC and 37ºC. The sensitivity of this agar to identify Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, Candida rugosa, Candida krusei and Candida parapsilosis ranged between 25 and 100% at 30ºC, 14% and 100% at 35ºC, 56% and 100% at 37ºC. The specificity of this agar was 100% at 30ºC, between 97% and 100% at 35ºC, 92% and 100% at 37ºC. The efficiency of this agar ranged between 88 and 100% at 30ºC, 83% and 100% at 35ºC, 88% and 100% at 37ºC. Each species also gave rise to a variety of colony colours ranging from pink to green to blue of different colony characteristics. Therefore, the chromogenic agar was found to be useful in our study for identifying clinical Candida isolates.

Matched MeSH terms: Culture Media/chemistry*
The Influence of Prior Modes of Growth, Temperature, Medium, and Substrate Surface on Biofilm Formation by Antibiotic-Resistant Campylobacter jejuni

Teh AH, Lee SM, Dykes GA

Curr Microbiol, 2016 Dec;73(6):859-866.
PMID: 27623781

Campylobacter jejuni is one of the most common causes of bacterial gastrointestinal food-borne infection worldwide. It has been suggested that biofilm formation may play a role in survival of these bacteria in the environment. In this study, the influence of prior modes of growth (planktonic or sessile), temperatures (37 and 42 °C), and nutrient conditions (nutrient broth and Mueller-Hinton broth) on biofilm formation by eight C. jejuni strains with different antibiotic resistance profiles was examined. The ability of these strains to form biofilm on different abiotic surfaces (stainless steel, glass, and polystyrene) as well as factors potentially associated with biofilm formation (bacterial surface hydrophobicity, auto-aggregation, and initial attachment) was also determined. The results showed that cells grown as sessile culture generally have a greater ability to form biofilm (P media, while growth at different temperatures affects biofilm formation in a strain-dependent manner. The strains were able to attach and form biofilms on different abiotic surfaces, but none of them demonstrated strong, complex, or structured biofilm formation. There were no clear trends between the bacterial surface hydrophobicity, auto-aggregation, attachment, and biofilm formation by the strains. This finding suggests that environmental factors did affect biofilm formation by C. jejuni, and they are more likely to persist in the environment in the form of mixed-species rather than monospecies biofilms.

Matched MeSH terms: Culture Media/chemistry
Generation of functional hepatocyte-like cells from human deciduous periodontal ligament stem cells

Vasanthan P, Jayaraman P, Kunasekaran W, Lawrence A, Gnanasegaran N, Govindasamy V, et al.

Naturwissenschaften, 2016 Aug;103(7-8):62.
PMID: 27379400 DOI: 10.1007/s00114-016-1387-7

Human deciduous periodontal ligament stem cells have been introduced for as an easily accessible source of stem cells from dental origin. Although recent studies have revealed the ability of these stem cells in multipotential attribute, their efficiency of hepatic lineage differentiation has not been addressed so far. The aim of this study is to investigate hepatic lineage fate competence of periodontal ligament stem cells through direct media induction. Differentiation of periodontal ligament stem cells into hepatocyte-like cells was conducted by the exposure of two phase media induction. First phase was performed in the presence of hepatocyte growth factors to induce a definitive endoderm formation. In the subsequent phase, the cells were treated with oncostatin M and dexamethosone followed by insulin and transferrin to generate hepatocyte-like cells. Hepatic-related characters of the generated hepatocyte-like cells were determined at both mRNA and protein level followed by functional assays. Foremost changes observed in the generation of hepatocyte-like cells were the morphological features in which these cells were transformed from fibroblastic shape to polygonal shape. Temporal expression of hepatic markers ranging from early endodermal up to late markers were detected in the hepatocyte-like cells. Crucial hepatic markers such as glycogen storage, albumin, and urea secretion were also shown. These findings exhibited the ability of periodontal ligament stem cells of dental origin to be directed into hepatic lineage fate. These cells can be regarded as an alternative autologous source in the usage of stem cell-based treatment for liver diseases.

Matched MeSH terms: Culture Media/chemistry
Fulltext Production of recombinant human epidermal growth factor in Pichia pastoris

Eissazadeh S, Moeini H, Dezfouli MG, Heidary S, Nelofer R, Abdullah MP

Braz J Microbiol, 2017 Apr-Jun;48(2):286-293.
PMID: 27998673 DOI: 10.1016/j.bjm.2016.10.017

This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27μg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.

Matched MeSH terms: Culture Media/chemistry
Determination of the Biological Efficiency and Antioxidant Potential of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Cultivated Using Different Agro-Wastes in Malaysia

Sudheer S, Alzorqi I, Ali A, Cheng PG, Siddiqui Y, Manickam S

Int J Med Mushrooms, 2018;20(1):89-100.
PMID: 29604916 DOI: 10.1615/IntJMedMushrooms.2017024588

This study investigates the cultivation of Ganoderma lucidum using different agricultural biomasses from Malaysia. Five different combinations of rubber wood sawdust, empty fruit bunch fiber, and mesocarp fiber from oil palm, alone and in combination, were used to cultivate G. lucidum. Although all the substrate combinations worked well to grow the mushroom, the highest biological efficiency was obtained from the combination of empty fruit bunch fiber with sawdust. A total yield of 27% was obtained from empty fruit bunch fiber with sawdust, followed by sawdust (26%), empty fruit bunch fiber (19%), mesocarp fiber with sawdust (19%), and mesocarp fiber (16%). The quality of mushrooms was proved by proximate analysis and detection of phenolic compounds and flavonoids. The antioxidant activity verified by DPPH, ferric-reducing ability of plasma, and ABTS analyses revealed that the empty fruit bunch fiber with sawdust had higher activity than the other substrates.

Matched MeSH terms: Culture Media/chemistry
Formulation of maize- and peanut-based semi-synthetic growth media for the ecophysiological studies of aflatoxigenic Aspergillus flavus in maize and peanut agro-ecosystems

Yazid SNE, Thanggavelu H, Mahror N, Selamat J, Samsudin NIP

Int J Food Microbiol, 2018 Oct 03;282:57-65.
PMID: 29913332 DOI: 10.1016/j.ijfoodmicro.2018.06.007

In studying the ecophysiology of fungal phytopathogens, several stages are involved (in vitro, greenhouse, in planta). Most in vitro studies extensively utilise the general growth media such as Potato Dextrose Agar and Malt Extract Agar. Although the crop components in these media serve as excellent carbon sources and yield luxuriant growth, they are not naturally contaminated with Aspergillus flavus and thus might result in under- or overestimation of its actual toxigenic potentials. Empirical data on the formulation of semi-synthetic growth medium mimicking the natural crop commonly contaminated by A. flavus for the ecophysiological studies in vitro are scarce. The present work was aimed at investigating the ecophysiology of A. flavus on commercial growth media (PDA, MEA); formulating maize- and peanut-based semi-synthetic growth media using two methods of raw material preparation (milling, hot water extraction) at different concentrations (1, 3, 5, 7, 9% w/v), and comparing the ecophysiological parameters between commercial and formulated growth media. Growth rates were obtained by computing the hyphal expansion data into y = mx + c equation. AFB1 was quantified using high performance liquid chromatography with fluorescence detector. Formulated media were found to yield significantly higher growth rates when compared to commercial media. However, commercial media yielded significantly higher AFB1 when compared to all formulated media. Between the two formulations, milling yielded significantly higher growth rates and AFB1 when compared to hot water extraction. Although in vitro data cannot directly extrapolate in planta performance, results obtained in the present work can be used to gauge the actual toxigenic potential of A. flavus in maize and peanut agro-ecosystems.

Matched MeSH terms: Culture Media/chemistry*
Fulltext Production and Characterization of a Bioflocculant Produced by Bacillussalmalaya 139SI-7 and Its Applications in Wastewater Treatment

Abu Tawila ZM, Ismail S, Dadrasnia A, Usman MM

Molecules, 2018 Oct 18;23(10).
PMID: 30340415 DOI: 10.3390/molecules23102689

The production, optimization, and characterization of the bioflocculant QZ-7 synthesized by a novel Bacillus salmalaya strain 139SI isolated from a private farm soil in Selangor, Malaysia, are reported. The flocculating activity of bioflocculant QZ-7 present in the selected strain was found to be 83.3%. The optimal culture for flocculant production was achieved after cultivation at 35.5 °C for 72 h at pH 7 ± 0.2, with an inoculum size of 5% (v/v) and sucrose and yeast extract as carbon and nitrogen sources. The maximum flocculating activity was found to be 92.6%. Chemical analysis revealed that the pure bioflocculant consisted of 79.08% carbohydrates and 15.4% proteins. The average molecular weight of the bioflocculant was calculated to be 5.13 × 10⁵ Da. Infrared spectrometric analysis showed the presence of carboxyl (COO-), hydroxyl (-OH), and amino (-NH₂) groups, polysaccharides and proteins. The bioflocculant QZ-7 exhibited a wide pH stability range from 4 to 7, with a flocculation activity of 85% at pH 7 ± 0.2. In addition, QZ-7 was thermally stable and retained more than 80% of its flocculating activity after being heated at 80 °C for 30 min. SEM analysis revealed that QZ-7 exhibited a clear crystalline brick-shaped structure. After treating wastewater, the bioflocculant QZ-7 showed significant flocculation performance with a COD removal efficiency of 93%, whereas a BOD removal efficiency of 92.4% was observed in the B. salmalaya strain 139SI. These values indicate the promising applications of the bioflocculant QZ-7 in wastewater treatment.

Matched MeSH terms: Culture Media/chemistry
Fulltext Biohydrogen Production by Antarctic Psychrotolerant Klebsiella sp. ABZ11

Mohammed A, Abdul-Wahab MF, Hashim M, Omar AH, Md Reba MN, Muhamad Said MF, et al.

Pol J Microbiol, 2018 11 20;67(3):283-290.
PMID: 30451444 DOI: 10.21307/pjm-2018-033

Lower temperature biohydrogen production has always been attractive, due to the lower energy requirements. However, the slow metabolic rate of psychrotolerant biohydrogen-producing bacteria is a common problem that affects their biohydrogen yield. This study reports on the improved substrate synthesis and biohydrogen productivity by the psychrotolerant Klebsiella sp. strain ABZ11, isolated from Antarctic seawater sample. The isolate was screened for biohydrogen production at 30°C, under facultative anaerobic condition. The isolate is able to ferment glucose, fructose and sucrose with biohydrogen production rate and yield of 0.8 mol/l/h and 3.8 mol/g, respectively at 10 g/l glucose concentration. It also showed 74% carbohydrate uptake and 95% oxygen uptake ability, and a wide growth temperature range with optimum at 37°C. Klebsiella sp. ABZ11 has a short biohydrogen production lag phase, fast substrate uptake and is able to tolerate the presence of oxygen in the culture medium. Thus, the isolate has a potential to be used for lower temperature biohydrogen production process.

Matched MeSH terms: Culture Media/chemistry
Exploring the additive bio-agent impacts upon ectoine production by Halomonas salina DSM5928T using corn steep liquor and soybean hydrolysate as nutrient supplement

Chen PW, Cui ZY, Ng HS, Chi-Wei Lan J

J Biosci Bioeng, 2020 Aug;130(2):195-199.
PMID: 32370929 DOI: 10.1016/j.jbiosc.2020.03.011

Ectoine production using inexpensive and renewable biomass resources has attracted great interest among the researchers due to the low yields of ectoine in current fermentation approaches that complicate the large-scale production of ectoine. In this study, ectoine was produced from corn steep liquor (CSL) and soybean hydrolysate (SH) in replacement to yeast extract as the nitrogen sources for the fermentation process. To enhance the bacterial growth and ectoine production, biotin was added to the Halomonas salina fermentation media. In addition, the effects addition of surfactants such as Tween 80 and saponin on the ectoine production were also investigated. Results showed that both the CSL and SH can be used as the nitrogen source substitutes in the fermentation media. Higher amount of ectoine (1781.9 mg L-1) was produced in shake flask culture with SH-containing media as compared to CSL-containing media. A total of 2537.0 mg L-1 of ectoine was produced at pH 7 when SH-containing media was applied in the 2 L batch fermentation. Moreover, highest amount of ectoine (1802.0 mg L-1) was recorded in the SH-containing shake flask culture with addition of 0.2 μm mL-1 biotin. This study demonstrated the efficacy of industrial waste as the nutrient supplement for the fermentation of ectoine production.

Matched MeSH terms: Culture Media/chemistry
A brief period of darkness induces changes in fatty acid biosynthesis towards accumulation of saturated fatty acids in Chlorella vulgaris UMT-M1 at stationary growth phase

Cha TS, Yee W, Phua PSP, Loh SH, Aziz A

Biotechnol Lett, 2021 Apr;43(4):803-812.
PMID: 33438120 DOI: 10.1007/s10529-021-03077-2

OBJECTIVE: The effects of a brief (3 days) and prolonged (6 days) period of incubation in darkness and light on the biomass content, lipid content and fatty acid profile in Chlorella vulgaris UMT-M1 were determined.
RESULTS: Three days of incubation in darkness increased saturated fatty acid (SFA) content from 34.0 to 41.4% but decreased monounsaturated fatty acid (MUFA) content from 36.7 to 29.8%. Palmitic acid (C16:0) content was increased from 23.2 to 28.9%, whereas oleic acid (C18:1) content was reduced from 35.4 to 28.8%. Total oil content was slightly decreased from 20.4 to 18.7% after 3 days of darkness, without a significant reduction in biomass compared to 3 days of incubation in light. Biomass and oil content was highest in cultures incubated for 6 days in light, however the stimulatory and inhibitory effects of darkness (or light) on SFA and MUFA content was no longer present at 6 days of incubation.
CONCLUSIONS: Findings from this study suggests that fatty acid composition in C. vulgaris could be modulated to favor either C16:0 or C18:1 by a brief period of either darkness or light incubation, prior to harvesting.

Matched MeSH terms: Culture Media/chemistry
Fulltext Limitation of nutrients stimulates musty odor production by Streptomyces sp. isolated from a tropical environment

Shudirman S, Abang Kassim A, Shamsol Anuar NS, Utsumi M, Shimizu K, Muhammad Yuzir MA, et al.

J Gen Appl Microbiol, 2021 Jul 31;67(3):92-99.
PMID: 33642451 DOI: 10.2323/jgam.2020.08.001

Musty odor production by actinomycetes is usually related to the presence of geosmin and 2-methylisoborneol (2-MIB), which are synthesized by enzymes encoded by the geoA and tpc genes, respectively. Streptomyces spp. strain S10, which was isolated from a water reservoir in Malaysia, has the ability to produce geosmin when cultivated in a basal salt (BS) solid medium, but no 2-MIB production occurred during growth in BS medium. Strain S10 could produce higher levels of geosmin when the phosphate concentration was limited to 0.05 mg/L, with a yield of 17.53 ± 3.12 ✕ 105 ng/L, compared with growth in BS medium. Interestingly, 2-MIB production was suddenly detected when the nitrate concentration was limited to 1.0 mg/L, with a yield of 1.4 ± 0.11 ✕ 105 ng/L. Therefore, it was concluded that phosphate- and nitrate-limiting conditions could induce the initial production of geosmin and 2-MIB by strain S10. Furthermore, a positive amplicon of geoA was detected in strain S10, but no tpc amplicon was detected by PCR analysis. Draft genome sequence analysis showed that one open reading frame (ORF) contained a conserved motif of geosmin synthase with 95% identity with geoA in Streptomyces coelicolor A3 (2). In the case of the tpc genes, it was found that one ORF showed 23% identity to the known tpc gene in S. coelicolor A3(2), but strain S10 lacked one motif in the N-terminus.

Matched MeSH terms: Culture Media/chemistry
Effects of simulated microgravity on gene expression and biological phenotypes of a single generation Caenorhabditis elegans cultured on 2 different media

Tee LF, Neoh HM, Then SM, Murad NA, Asillam MF, Hashim MH, et al.

Life Sci Space Res (Amst), 2017 Nov;15:11-17.
PMID: 29198309 DOI: 10.1016/j.lssr.2017.06.002

Studies of multigenerational Caenorhabditis elegans exposed to long-term spaceflight have revealed expression changes of genes involved in longevity, DNA repair, and locomotion. However, results from spaceflight experiments are difficult to reproduce as space missions are costly and opportunities are rather limited for researchers. In addition, multigenerational cultures of C. elegans used in previous studies contribute to mixture of gene expression profiles from both larvae and adult worms, which were recently reported to be different. Usage of different culture media during microgravity simulation experiments might also give rise to differences in the gene expression and biological phenotypes of the worms. In this study, we investigated the effects of simulated microgravity on the gene expression and biological phenotype profiles of a single generation of C. elegans worms cultured on 2 different culture media. A desktop Random Positioning Machine (RPM) was used to simulate microgravity on the worms for approximately 52 to 54 h. Gene expression profile was analysed using the Affymetrix GeneChip® C. elegans 1.0 ST Array. Only one gene (R01H2.2) was found to be downregulated in nematode growth medium (NGM)-cultured worms exposed to simulated microgravity. On the other hand, eight genes were differentially expressed for C. elegans Maintenance Medium (CeMM)-cultured worms in microgravity; six were upregulated, while two were downregulated. Five of the upregulated genes (C07E3.15, C34H3.21, C32D5.16, F35H8.9 and C34F11.17) encode non-coding RNAs. In terms of biological phenotype, we observed that microgravity-simulated worms experienced minimal changes in terms of lifespan, locomotion and reproductive capabilities in comparison with the ground controls. Taking it all together, simulated microgravity on a single generation of C. elegans did not confer major changes to their gene expression and biological phenotype. Nevertheless, exposure of the worms to microgravity lead to higher expression of non-coding RNA genes, which may play an epigenetic role in the worms during longer terms of microgravity exposure.

Matched MeSH terms: Culture Media/chemistry*
Fulltext A refined medium to enhance the antimicrobial activity of postbiotic produced by Lactiplantibacillus plantarum RS5

Ooi MF, Foo HL, Loh TC, Mohamad R, Rahim RA, Ariff A

Sci Rep, 2021 Apr 07;11(1):7617.
PMID: 33828119 DOI: 10.1038/s41598-021-87081-6

Postbiotic RS5, produced by Lactiplantibacillus plantarum RS5, has been identified as a promising alternative feed supplement for various livestock. This study aimed to lower the production cost by enhancing the antimicrobial activity of the postbiotic RS5 by improving the culture density of L. plantarum RS5 and reducing the cost of growth medium. A combination of conventional and statistical-based approaches (Fractional Factorial Design and Central Composite Design of Response Surface Methodology) was employed to develop a refined medium for the enhancement of the antimicrobial activity of postbiotic RS5. A refined medium containing 20 g/L of glucose, 27.84 g/L of yeast extract, 5.75 g/L of sodium acetate, 1.12 g/L of Tween 80 and 0.05 g/L of manganese sulphate enhanced the antimicrobial activity of postbiotic RS5 by 108%. The cost of the production medium was reduced by 85% as compared to the commercially available de Man, Rogosa and Sharpe medium that is typically used for Lactobacillus cultivation. Hence, the refined medium has made the postbiotic RS5 more feasible and cost-effective to be adopted as a feed supplement for various livestock industries.

Matched MeSH terms: Culture Media/chemistry*
Degradation and transformation of anthracene by white-rot fungus Armillaria sp. F022

Hadibarata T, Zubir MM, Rubiyatno, Chuang TZ, Yusoff AR, Salim MR, et al.

Folia Microbiol (Praha), 2013 Sep;58(5):385-91.
PMID: 23307571 DOI: 10.1007/s12223-013-0221-2

Characterization of anthracene metabolites produced by Armillaria sp. F022 was performed in the enzymatic system. The fungal culture was conducted in 100-mL Erlenmeyer flask containing mineral salt broth medium (20 mL) and incubated at 120 rpm for 5-30 days. The culture broth was then centrifuged at 10,000 rpm for 45 min to obtain the extract. Additionally, the effect of glucose consumption, laccase activity, and biomass production in degradation of anthracene were also investigated. Approximately, 92 % of the initial concentration of anthracene was degraded within 30 days of incubation. Dynamic pattern of the biomass production was affected the laccase activity during the experiment. The biomass of the fungus increased with the increasing of laccase activity. The isolation and characterization of four metabolites indicated that the structure of anthracene was transformed by Armillaria sp. F022 in two routes. First, anthracene was oxidized to form anthraquinone, benzoic acid, and second, converted into other products, 2-hydroxy-3-naphthoic acid and coumarin. Gas chromatography-mass spectrometry analysis also revealed that the molecular structure of anthracene was transformed by the action of the enzyme, generating a series of intermediate compounds such as anthraquinone by ring-cleavage reactions. The ligninolytic enzymes expecially free extracellular laccase played an important role in the transformation of anthracene during degradation period.

Matched MeSH terms: Culture Media/chemistry
Applications of the Box-Wilson design model for bio-hydrogen production using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564)

Alalayah WM, Kalil MS, Kadhum AA, Jahim J, Zaharim A, Alauj NM, et al.

Pak J Biol Sci, 2010 Jul 15;13(14):674-82.
PMID: 21848059

Box-Wilson design (BWD) model was applied to determine the optimum values of influencing parameters in anaerobic fermentation to produce hydrogen using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). The main focus of the study was to find the optimal relationship between the hydrogen yield and three variables including initial substrate concentration, initial medium pH and reaction temperature. Microbial growth kinetic parameters for hydrogen production under anaerobic conditions were determined using the Monod model with incorporation of a substrate inhibition term. The values of micro(max) (maximum specific growth rate) and K, (saturation constant) were 0.398 h(-1) and 5.509 g L(-1), respectively, using glucose as the substrate. The experimental substrate and biomass-concentration profiles were in good agreement with those obtained by the kinetic-model predictions. By varying the conditions of the initial substrate concentration (1-40 g L(-1)), reaction temperature (25-40 degrees C) and initial medium pH (4-8), the model predicted a maximum hydrogen yield of 3.24 mol H2 (mol glucose)(-1). The experimental data collected utilising this design was successfully fitted to a second-order polynomial model. An optimum operating condition of 10 g L(-1) initial substrate concentration, 37 degrees C reaction temperature and 6.0 +/- 0.2 initial medium pH gave 80% of the predicted maximum yield of hydrogen where as the experimental yield obtained in this study was 77.75% exhibiting a close accuracy between estimated and experimental values. This is the first report to predict bio-hydrogen yield by applying Box-Wilson Design in anaerobic fermentation while optimizing the effects of environmental factors prevailing there by investigating the effects of environmental factors.

Matched MeSH terms: Culture Media/chemistry
Fulltext Optimized medium via statistical approach enhanced threonine production by Pediococcus pentosaceus TL-3 isolated from Malaysian food

Lim YH, Foo HL, Loh TC, Mohamad R, Abdul Rahim R, Idrus Z

Microb Cell Fact, 2019 Jul 22;18(1):125.
PMID: 31331395 DOI: 10.1186/s12934-019-1173-2

BACKGROUND: Threonine is an essential amino acid that is extensively used in livestock industry as feed supplement due to its pronounced effect in improving the growth performance of animals. Application of genetically engineered bacteria for amino acid production has its share of controversies after eosinophils myalgia syndrome outbreak in 1980s. This has urged for continuous search for a food grade producer as a safer alternative for industrial amino acid production. Lactic acid bacteria (LAB) appear as an exceptional candidate owing to their non-pathogenic nature and reputation of Generally Recognized as Safe (GRAS) status. Recently, we have identified a LAB, Pediococcus pentosaceus TL-3, isolated from Malaysian food as a potential threonine producer. Thus, the objective of this study was to enhance the threonine production by P. pentosaceus TL-3 via optimized medium developed by using Plackett-Burman design (PBD) and central composite design (CCD).
RESULTS: Molasses, meat extract, (NH4)2SO4, and MnSO4 were identified as the main medium components for threonine production by P. pentosaceus TL-3. The optimum concentration of molasses, meat extract, (NH4)2SO4 and MnSO4 were found to be 30.79 g/L, 25.30 g/L, 8.59 g/L, and 0.098 g/L respectively based on model obtained in CCD with a predicted net threonine production of 123.07 mg/L. The net threonine production by P. pentosaceus TL-3 in the optimized medium was enhanced approximately 2 folds compared to the control.
CONCLUSIONS: This study has revealed the potential of P. pentosaceus TL-3 as a safer alternative to produce threonine. Additionally, the current study has identified the key medium components affecting the production of threonine by P. pentosaceus TL-3, followed by optimization of their concentrations by means of statistical approach. The findings of this study could act as a guideline for the future exploration of amino acid production by LAB.

Matched MeSH terms: Culture Media/chemistry*
Production of single cell oil by Lipomyces starkeyi from waste plant oil generated by the palm oil mill industry

Zain NA, Kahar P, Sudesh K, Ogino C, Kondo A

J Biosci Bioeng, 2024 Aug;138(2):153-162.
PMID: 38777650 DOI: 10.1016/j.jbiosc.2024.04.005

Only a few reports available about the assimilation of hydrophobic or oil-based feedstock as carbon sources by Lipomyces starkeyi. In this study, the ability of L. starkeyi to efficiently utilize free fatty acids (FFAs) and real biomass like palm acid oil (PAO) as well as crude palm kernel oil (CPKO) for growth and lipid production was investigated. PAO, CPKO, and FFAs were evaluated as sole carbon sources or in the mixed medium containing glucose. L. starkeyi was able to grow on the medium supplemented with PAO and FFAs, which contained long-chain length FAs and accumulated lipids up to 35% (w/w) of its dry cell weight. The highest lipid content and lipid concentration were achieved at 50% (w/w) and 10.1 g/L, respectively, when L. starkeyi was cultured in nitrogen-limited mineral medium (-NMM) supplemented with PAO emulsion. Hydrophobic substrate like PAO could be served as promising carbon source for L. starkeyi.

Matched MeSH terms: Culture Media/chemistry
Extracellular BSA-degrading SAPs in the rare pathogen Meyerozyma guilliermondii strain SO as potential virulence factors in candidiasis

Lim SJ, Noor NDM, Sabri S, Ali MSM, Salleh AB, Oslan SN

Microb Pathog, 2024 Aug;193:106773.
PMID: 38960213 DOI: 10.1016/j.micpath.2024.106773

Meyerozyma guilliermondii (Candida guilliermondii) is one of the Candida species associated with invasive candidiasis. With the potential for expressing industrially important enzymes, M. guilliermondii strain SO possessed 99 % proteome similarity with the clinical ATCC 6260 isolate and showed pathogenicity towards zebrafish embryos. Recently, three secreted aspartyl proteinases (SAPs) were computationally identified as potential virulence factors in this strain without in vitro verification of SAP activity. The quantification of Candida SAPs activity in liquid broth were also scarcely reported. Thus, this study aimed to characterize M. guilliermondii strain SO's ability to produce SAPs (MgSAPs) in different conditions (morphology and medium) besides analyzing its growth profile. MgSAPs' capability to cleave bovine serum albumin (BSA) was also determined to propose that MgSAPs as the potential virulence factors compared to the avirulent Saccharomyces cerevisiae. M. guilliermondii strain SO produced more SAPs (higher activity) in yeast nitrogen base-BSA-dextrose broth compared to yeast extract-BSA-dextrose broth despite insignificantly different SAP activity in both planktonic and biofilm cells. FeCl3 supplementation significantly increased the specific protein activity (∼40 %). The BSA cleavage by MgSAPs at an acidic pH was proven through semi-quantitative SDS-PAGE, sharing similar profile with HIV-1 retropepsin. The presented work highlighted the MgSAPs on fungal cell wall and extracellular milieu during host infection could be corroborated to the quantitative production in different growth modes presented herein besides shedding lights on the potential usage of retropepsin's inhibitors in treating candidiasis. Molecular and expression analyses of MgSAPs and their deletion should be further explored to attribute their respective virulence effects.

Matched MeSH terms: Culture Media/chemistry
Fulltext Quantification of mesenchymal stem cell growth rates through secretory and excretory biomolecules in conditioned media via Fresnel reflection

Ahmad H, Thambiratnam K, Zulkifli AZ, Lawrence A, Jasim AA, Kunasekaran W, et al.

Sensors (Basel), 2013 Sep 30;13(10):13276-88.
PMID: 24084118 DOI: 10.3390/s131013276

An efficient and low cost optical method for directly measuring the concentration of homogenous biological solutes is proposed and demonstrated. The proposed system operates by Fresnel reflection, with a flat-cleaved single-mode fiber serving as the sensor probe. A laser provides a 12.9 dBm sensor signal at 1,550 nm, while a computer-controlled optical power meter measures the power of the signal returned by the probe. Three different mesenchymal stem cell (MSC) lines were obtained, sub-cultured and trypsinized daily over 9 days. Counts were measured using a haemocytometer and the conditioned media (CM) was collected daily and stored at -80 °C. MSCs release excretory biomolecules proportional to their growth rate into the CM, which changes the refractive index of the latter. The sensor is capable of detecting changes in the number of stem cells via correlation to the change in the refractive index of the CM, with the measured power loss decreasing approximately 0.4 dB in the CM sample per average 1,000 cells in the MSC subculture. The proposed system is highly cost-effective, simple to deploy, operate, and maintain, is non-destructive, and allows reliable real-time measurement of various stem cell proliferation parameters.

Matched MeSH terms: Culture Media/chemistry
Enhancement of anti-candidal activity of endophytic fungus Phomopsis sp. ED2, isolated from Orthosiphon stamineus Benth, by incorporation of host plant extract in culture medium

Yenn TW, Lee CC, Ibrahim D, Zakaria L

J Microbiol, 2012 Aug;50(4):581-5.
PMID: 22923105 DOI: 10.1007/s12275-012-2083-8

This study examined the effect of host extract in the culture medium on anti-candidal activity of Phomopsis sp. ED2, previously isolated from the medicinal herb Orthosiphon stamineus Benth. Interestingly, upon addition of aqueous host extract to the culture medium, the ethyl acetate extract prepared from fermentative broth exhibited moderate anti-candidal activity in a disc diffusion assay. The minimal inhibitory concentration of this extract was 62.5 μg/ml and it only exhibited fungistatic activity against C. albicans. In the time-kill study, a 50% growth reduction of C. albicans was observed at 31.4 h for extract from the culture incorporating host extract. In the bioautography assay, only one single spot (Rf 0.59) developed from the extract exhibited anti-candidal activity. A spot with the a similar Rf was not detected for the crude extract from YES broth without host extract. This indicated that the terpenoid anti-candidal compound was only produced when the host extract was introduced into the medium. The study concluded that the incorporation of aqueous extract of the host plant into the culture medium significantly enhanced the anti-candidal activity of Phomopsis sp. ED2.

Matched MeSH terms: Culture Media/chemistry*

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