Willy Yee - Academia.edu (original) (raw)

Papers by Willy Yee

Journal of sustainability science and management, 2012

Molecules

Microalgae have become a popular area of research over the past few decades due to their enormous... more Microalgae have become a popular area of research over the past few decades due to their enormous benefits to various sectors, such as pharmaceuticals, biofuels, and food and feed. Nevertheless, the benefits of microalgae cannot be fully exploited without the optimization of their upstream production. The growth of microalgae is commonly measured based on the optical density of the sample. However, the presence of debris in the culture and the optical absorption of the intercellular components affect the accuracy of this measurement. As a solution, this paper introduces the direct optical detection of glucose molecules at 940–960 nm to accurately measure the growth of microalgae. In addition, this paper also discusses the effects of the presence of glucose on the absorption of free water molecules in the culture. The potential of the optical detection of glucose as a complement to the commonly used optical density measurement at 680 nm is discussed in this paper. Lastly, a few recom...

Indian Journal of Microbiology, Dec 14, 2017

2-mercaptoethanol (2-ME), alongside polyvinylpyrrolidone is commonly used in plant DNA extraction... more 2-mercaptoethanol (2-ME), alongside polyvinylpyrrolidone is commonly used in plant DNA extractions to deal with polyphenols, which could interfere with extraction and downstream applications. 2-ME is also commonly used to denature proteins and nucleases, especially RNAses. On the contrary, we found that the presence of 2-ME in lysis buffer interfered with DNA extraction from 12 strains of freshwater microalgae, resulting in DNA with poor integrity. We also found that the TNES-urea buffer, commonly used for preservation and DNA extraction from fish, appears as effective as the SDS and CTAB buffer for some microalgae strains. Results from our study suggests that the inclusion of 2-ME in DNA extraction protocols may be detrimental for isolation of good quality DNA from freshwater microalgae, and therefore recommend eliminating it or testing varying concentrations of 2-ME when developing species-specific extraction protocols for microalgae. Keywords 2-Mercaptoethanol Á DNA extraction Á Microalgae Á Lysis buffer Á TNES-urea Electronic supplementary material The online version of this article (

3 Biotech, 2018

In this work, a simple and inexpensive physical lysis method using a cordless drill fitted with a... more In this work, a simple and inexpensive physical lysis method using a cordless drill fitted with a plastic pellet pestle and 150 mg of sterile sea sand was established for the extraction of DNA from six strains of freshwater microalgae. This lysis method was also tested for RNA extraction from two microalgal strains. Lysis duration between 15 and 120 s using the cetyltrimethyl ammonium bromide (CTAB) buffer significantly increased the yield of DNA from four microalgalstrains ( NS16, sp. NS6, sp. DPBC1 and sp. DPBB10) compared to control. It was also found that grinding was not required to obtain DNA from two strains of microalgae ( sp. NPA14 and sp. BM3). The average DNA yield obtained using this lysis method was between 62.5 and 78.9 ng/mg for NS16, 42.2-247.0 ng/mg for sp. NS6, 70.2-110.9 ng/mg for sp. DPBC1 and 142.8-164.8 ng/mg for sp. DPBB10. DNA obtained using this method was sufficiently pure for PCR amplification. Extraction of total RNA from NS16 and sp. NPD7 using this lysi...

World Journal of Microbiology and Biotechnology, 2016

Over the years, microalgae have been identified to be a potential source of commercially importan... more Over the years, microalgae have been identified to be a potential source of commercially important products such as pigments, polysaccharides, polyunsaturated fatty acids and in particular, biofuels. Current demands for sustainable fuel sources and bioproducts has led to an extensive search for promising strains of microalgae for large scale cultivation. Prospective strains identified for these purposes were among others, mainly from the genera Hematococcus, Dunaliella, Botryococcus, Chlorella, Scenedesmus and Nannochloropsis. Recently, microalgae from the Selenastraceae emerged as potential candidates for biodiesel production. Strains from the Selenastraceae such as Monoraphidium sp. FXY-10, M. contortum SAG 47.80, Ankistrodesmus sp. SP2-15 and M. minutum were high biomass and lipid producers when cultivated under optimal conditions. A number of Selenastraceae strains were also reported to be suitable for cultivation in wastewater. This review highlights recent reports on potential strains from the Selenastraceae for biodiesel production and contrasts their biomass productivity, lipid productivity as well as fatty acid profile. Cultivation strategies employed to enhance their biomass and lipid productivity as well as to reduce feedstock cost are also discussed in this paper.

Bioresource Technology, 2015

In order to assess the feasibility of various carbon sources and plant materials in increasing th... more In order to assess the feasibility of various carbon sources and plant materials in increasing the growth rate and biomass productivity of Monoraphidium griffithii, ten carbon sources as well as six plant materials were tested in mixotrophic cultures with or without aeration. It was found that glucose, fructose, maltose, sodium acetate and mannitol were potential carbon sources for growth enhancement of M. griffithii. Supplementation of culture medium with these carbon sources resulted in approximately 1-4-fold increase in cell density compared to control in a small scale culture. In a larger scale mixotrophic culture with aeration, 0.05% mannitol and 0.1% fructose resulted in a decent 1-1.5-fold increase in final cell density, approximately 2-fold increase in growth rate and 0.5-1-fold increase in dry biomass weight. Findings from this study suggests that glucose, fructose, maltose and mannitol were potential organic carbon sources for mixotrophic culture of M. griffithii.

World Journal of Microbiology and Biotechnology, 2011

The successful establishment of an Agrobacterium-mediated transformation method and optimisation ... more The successful establishment of an Agrobacterium-mediated transformation method and optimisation of six critical parameters known to influence the efficacy of Agrobacterium T-DNA transfer in the unicellular microalga Chlorella vulgaris (UMT-M1) are reported. Agrobacterium tumefaciens strain LBA4404 harbouring the binary vector pCAMBIA1304 containing the gfp:gusA fusion reporter and a hygromycin phosphotransferase (hpt) selectable marker driven by the CaMV35S promoter were used for transformation. Transformation frequency was assessed by monitoring transient b-glucuronidase (GUS) expression 2 days post-infection. It was found that cocultivation temperature at 24°C, co-cultivation medium at pH 5.5, 3 days of co-cultivation, 150 lM acetosyringone, Agrobacterium density of 1.0 units (OD 600) and 2 days of pre-culture were optimum variables which produced the highest number of GUS-positive cells (8.8-20.1%) when each of these parameters was optimised individually. Transformation conducted with the combination of all optimal parameters above produced 25.0% of GUS-positive cells, which was almost a threefold increase from 8.9% obtained from un-optimised parameters. Evidence of transformation was further confirmed in 30% of 30 randomly-selected hygromycin B (20 mg L-1) resistant colonies by polymerase chain reaction (PCR) using gfp:gusA and hpt-specific primers. The developed transformation method is expected to facilitate the genetic improvement of this commercially-important microalga.

Journal of Microbiological Methods, 2011

The use of acetosyringone in Agrobacterium-mediated gene transfer into plant hosts has been favor... more The use of acetosyringone in Agrobacterium-mediated gene transfer into plant hosts has been favored for the past few decades. The influence of other phenolic compounds and their effectiveness in Agrobacterium-mediated plant transformation systems has been neglected. In this study, the efficacy of four phenolic compounds on Agrobacterium-mediated transformation of the unicellular green alga Nannochloropsis sp. (Strain UMT-M3) was assessed by using β-glucuronidase (GUS) assay. We found that cinnamic acid, vanillin and coumarin produced higher percentages of GUS positive cells as compared to acetosyringone. These results also show that the presence of methoxy group in the phenolic compounds may not be necessary for Agrobacterium vir gene induction and receptor binding as suggested by previous studies. These findings provide possible alternative Agrobacterium vir gene inducers that are more potent as compared to the commonly used acetosyringone in achieving high efficiency of Agrobacterium-mediated transformation in microalgae and possibly for other plants.

World Journal of Microbiology and Biotechnology, 2022

Microalgae are photosynthetic organisms functioning as the green bio-factories for various pharma... more Microalgae are photosynthetic organisms functioning as the green bio-factories for various pharmaceutical and biofuel products. To date, numerous attempts have been carried out to manipulate culture conditions to maximize the production of the desired metabolites. Because light is the energy source of microalgae for their growth and metabolites biosynthesis, it has been one of the most investigated variables emphasized on the deep understanding of how microalgae respond towards light changes as an external stimulus. This review discusses the effects of different light sources, light intensities, light wavelengths and length of photoperiod on various microalgae species, especially in terms of biomass and lipid productivity. Additionally, the relationship between photoregulation processes and lipid productivity of microalgae are also deliberated. The current available approaches of microalgae mass cultivation, including different types of open and closed systems are recapitulated with the intention to highlight the significant insights for the design of future photoreactors.

World Journal of Microbiology and Biotechnology

Applied Biochemistry and Biotechnology

Journal of sustainability science and management, 2012

Molecules

Microalgae have become a popular area of research over the past few decades due to their enormous... more Microalgae have become a popular area of research over the past few decades due to their enormous benefits to various sectors, such as pharmaceuticals, biofuels, and food and feed. Nevertheless, the benefits of microalgae cannot be fully exploited without the optimization of their upstream production. The growth of microalgae is commonly measured based on the optical density of the sample. However, the presence of debris in the culture and the optical absorption of the intercellular components affect the accuracy of this measurement. As a solution, this paper introduces the direct optical detection of glucose molecules at 940–960 nm to accurately measure the growth of microalgae. In addition, this paper also discusses the effects of the presence of glucose on the absorption of free water molecules in the culture. The potential of the optical detection of glucose as a complement to the commonly used optical density measurement at 680 nm is discussed in this paper. Lastly, a few recom...

Indian Journal of Microbiology, Dec 14, 2017

2-mercaptoethanol (2-ME), alongside polyvinylpyrrolidone is commonly used in plant DNA extraction... more 2-mercaptoethanol (2-ME), alongside polyvinylpyrrolidone is commonly used in plant DNA extractions to deal with polyphenols, which could interfere with extraction and downstream applications. 2-ME is also commonly used to denature proteins and nucleases, especially RNAses. On the contrary, we found that the presence of 2-ME in lysis buffer interfered with DNA extraction from 12 strains of freshwater microalgae, resulting in DNA with poor integrity. We also found that the TNES-urea buffer, commonly used for preservation and DNA extraction from fish, appears as effective as the SDS and CTAB buffer for some microalgae strains. Results from our study suggests that the inclusion of 2-ME in DNA extraction protocols may be detrimental for isolation of good quality DNA from freshwater microalgae, and therefore recommend eliminating it or testing varying concentrations of 2-ME when developing species-specific extraction protocols for microalgae. Keywords 2-Mercaptoethanol Á DNA extraction Á Microalgae Á Lysis buffer Á TNES-urea Electronic supplementary material The online version of this article (

3 Biotech, 2018

In this work, a simple and inexpensive physical lysis method using a cordless drill fitted with a... more In this work, a simple and inexpensive physical lysis method using a cordless drill fitted with a plastic pellet pestle and 150 mg of sterile sea sand was established for the extraction of DNA from six strains of freshwater microalgae. This lysis method was also tested for RNA extraction from two microalgal strains. Lysis duration between 15 and 120 s using the cetyltrimethyl ammonium bromide (CTAB) buffer significantly increased the yield of DNA from four microalgalstrains ( NS16, sp. NS6, sp. DPBC1 and sp. DPBB10) compared to control. It was also found that grinding was not required to obtain DNA from two strains of microalgae ( sp. NPA14 and sp. BM3). The average DNA yield obtained using this lysis method was between 62.5 and 78.9 ng/mg for NS16, 42.2-247.0 ng/mg for sp. NS6, 70.2-110.9 ng/mg for sp. DPBC1 and 142.8-164.8 ng/mg for sp. DPBB10. DNA obtained using this method was sufficiently pure for PCR amplification. Extraction of total RNA from NS16 and sp. NPD7 using this lysi...

World Journal of Microbiology and Biotechnology, 2016

Over the years, microalgae have been identified to be a potential source of commercially importan... more Over the years, microalgae have been identified to be a potential source of commercially important products such as pigments, polysaccharides, polyunsaturated fatty acids and in particular, biofuels. Current demands for sustainable fuel sources and bioproducts has led to an extensive search for promising strains of microalgae for large scale cultivation. Prospective strains identified for these purposes were among others, mainly from the genera Hematococcus, Dunaliella, Botryococcus, Chlorella, Scenedesmus and Nannochloropsis. Recently, microalgae from the Selenastraceae emerged as potential candidates for biodiesel production. Strains from the Selenastraceae such as Monoraphidium sp. FXY-10, M. contortum SAG 47.80, Ankistrodesmus sp. SP2-15 and M. minutum were high biomass and lipid producers when cultivated under optimal conditions. A number of Selenastraceae strains were also reported to be suitable for cultivation in wastewater. This review highlights recent reports on potential strains from the Selenastraceae for biodiesel production and contrasts their biomass productivity, lipid productivity as well as fatty acid profile. Cultivation strategies employed to enhance their biomass and lipid productivity as well as to reduce feedstock cost are also discussed in this paper.

Bioresource Technology, 2015

In order to assess the feasibility of various carbon sources and plant materials in increasing th... more In order to assess the feasibility of various carbon sources and plant materials in increasing the growth rate and biomass productivity of Monoraphidium griffithii, ten carbon sources as well as six plant materials were tested in mixotrophic cultures with or without aeration. It was found that glucose, fructose, maltose, sodium acetate and mannitol were potential carbon sources for growth enhancement of M. griffithii. Supplementation of culture medium with these carbon sources resulted in approximately 1-4-fold increase in cell density compared to control in a small scale culture. In a larger scale mixotrophic culture with aeration, 0.05% mannitol and 0.1% fructose resulted in a decent 1-1.5-fold increase in final cell density, approximately 2-fold increase in growth rate and 0.5-1-fold increase in dry biomass weight. Findings from this study suggests that glucose, fructose, maltose and mannitol were potential organic carbon sources for mixotrophic culture of M. griffithii.

World Journal of Microbiology and Biotechnology, 2011

The successful establishment of an Agrobacterium-mediated transformation method and optimisation ... more The successful establishment of an Agrobacterium-mediated transformation method and optimisation of six critical parameters known to influence the efficacy of Agrobacterium T-DNA transfer in the unicellular microalga Chlorella vulgaris (UMT-M1) are reported. Agrobacterium tumefaciens strain LBA4404 harbouring the binary vector pCAMBIA1304 containing the gfp:gusA fusion reporter and a hygromycin phosphotransferase (hpt) selectable marker driven by the CaMV35S promoter were used for transformation. Transformation frequency was assessed by monitoring transient b-glucuronidase (GUS) expression 2 days post-infection. It was found that cocultivation temperature at 24°C, co-cultivation medium at pH 5.5, 3 days of co-cultivation, 150 lM acetosyringone, Agrobacterium density of 1.0 units (OD 600) and 2 days of pre-culture were optimum variables which produced the highest number of GUS-positive cells (8.8-20.1%) when each of these parameters was optimised individually. Transformation conducted with the combination of all optimal parameters above produced 25.0% of GUS-positive cells, which was almost a threefold increase from 8.9% obtained from un-optimised parameters. Evidence of transformation was further confirmed in 30% of 30 randomly-selected hygromycin B (20 mg L-1) resistant colonies by polymerase chain reaction (PCR) using gfp:gusA and hpt-specific primers. The developed transformation method is expected to facilitate the genetic improvement of this commercially-important microalga.

Journal of Microbiological Methods, 2011

The use of acetosyringone in Agrobacterium-mediated gene transfer into plant hosts has been favor... more The use of acetosyringone in Agrobacterium-mediated gene transfer into plant hosts has been favored for the past few decades. The influence of other phenolic compounds and their effectiveness in Agrobacterium-mediated plant transformation systems has been neglected. In this study, the efficacy of four phenolic compounds on Agrobacterium-mediated transformation of the unicellular green alga Nannochloropsis sp. (Strain UMT-M3) was assessed by using β-glucuronidase (GUS) assay. We found that cinnamic acid, vanillin and coumarin produced higher percentages of GUS positive cells as compared to acetosyringone. These results also show that the presence of methoxy group in the phenolic compounds may not be necessary for Agrobacterium vir gene induction and receptor binding as suggested by previous studies. These findings provide possible alternative Agrobacterium vir gene inducers that are more potent as compared to the commonly used acetosyringone in achieving high efficiency of Agrobacterium-mediated transformation in microalgae and possibly for other plants.

World Journal of Microbiology and Biotechnology, 2022

Microalgae are photosynthetic organisms functioning as the green bio-factories for various pharma... more Microalgae are photosynthetic organisms functioning as the green bio-factories for various pharmaceutical and biofuel products. To date, numerous attempts have been carried out to manipulate culture conditions to maximize the production of the desired metabolites. Because light is the energy source of microalgae for their growth and metabolites biosynthesis, it has been one of the most investigated variables emphasized on the deep understanding of how microalgae respond towards light changes as an external stimulus. This review discusses the effects of different light sources, light intensities, light wavelengths and length of photoperiod on various microalgae species, especially in terms of biomass and lipid productivity. Additionally, the relationship between photoregulation processes and lipid productivity of microalgae are also deliberated. The current available approaches of microalgae mass cultivation, including different types of open and closed systems are recapitulated with the intention to highlight the significant insights for the design of future photoreactors.

World Journal of Microbiology and Biotechnology

Applied Biochemistry and Biotechnology