piroonporn srimongkol | Chulalongkorn University (original) (raw)

Papers by piroonporn srimongkol

PLOS ONE, Sep 2, 2021

When fish are processed, fish bone becomes a key component of the waste, but to date very few res... more When fish are processed, fish bone becomes a key component of the waste, but to date very few researchers have sought to use fish bone to prepare protein hydrolysates as a means of adding value to the final product. This study, therefore, examines the potential of salmon bone, through an analysis of the benefits of its constituent components, namely fat, moisture, protein, and ash. In particular, the study seeks to optimize the process of enzymatic hydrolysis of salmon bone with trypsin in order to produce angiotensin-I converting enzyme (ACE) inhibitory peptides making use of response surface methodology in combination with central composite design (CCD). Optimum hydrolysis conditions concerning DH (degree of hydrolysis) and ACE-inhibitory activity were initially determined using the response surface model. Having thus determined which of the salmon bone protein hydrolysates (SBPH) offered the greatest level of ACE-inhibitory activity, these SBPH were duly selected to undergo ultrafiltration for further fractionation. It was found that the greatest ACE-inhibitory activity was achieved by the SBPH fraction which had a molecular weight lower than 0.65 kDa. This fraction underwent further purification using RP-HPLC, revealing that the F 7 fraction offered the best ACE-inhibitory activity. For ACE inhibition, the ideal peptide in the context of the F 7 fraction comprised eight amino acids: Phe-Cys-Leu-Tyr-Glu-Leu-Ala-Arg (FCLYELAR), while analysis of the Lineweaver-Burk plot revealed that the FCLYELAR peptide can serve as an uncompetitive ACE inhibitor. An examination of the molecular docking process showed that the FCLYELAR peptide was primarily able to provide ACE-inhibitory qualities as a consequence of the hydrogen bond interactions taking place between ACE and the peptide. Furthermore, upon isolation form the SBPH, the ACEinhibitory peptide demonstrated ACE-inhibitory capabilities in vitro, underlining its potential for applications in the food and pharmaceutical sectors.

RSC Advances, 2021

De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry. DORB ... more De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry. DORB may prove to be an important protein source, and also contains many other micronutrients. This study has the principal aim of optimizing the process of DORB protein hydrolysate preparation, and then testing the hydrolysate to determine its lipase inhibitory activity. DORB underwent hydrolysis using Alcalase® and response surface methodology (RSM). The resulting degree of hydrolysis (DH) was then monitored along with the extent of any lipase inhibitory activity. The optimum levels of lipase inhibition were obtained at a temperature of 49.88 C, a duration of 150.43 minutes, and 1.53% Alcalase® used for the sample 5% (w/v) solution. In these conditions, the DH value was 35.65%, and the IC 50 value for lipase inhibitory activity was 2.84 mg mL À1. Five ranges of different molecular weights were obtained via fractionation, whereupon it was determined that the highest level of inhibitory activity was achieved by the <0.65 kDa fraction. This fraction was then further purified via RP-HPLC, and the resulting peak had a retention time of 21.75 minutes (F 2 sub-fraction) and exhibited high lipase inhibitory activity. Mass spectrometry was used to determine the amino acid sequence for this peak, identified as FYLGYCDY. This particular peptide is categorized as bitter, with a non-toxic profile, and having poor water solubility. The synthesized form of this peptide showed lipase inhibitory activity measured by an IC 50 value of 0.47 AE 0.02 mM. The Lineweaver-Burk plot revealed that FYLGYCDY is a non-competitive inhibitor, while analysis of the docking results provided details of the FYLGYCDY peptide binding site with the porcine pancreatic lipase (PPL) complex, which is a competitive type. It can be inferred from these findings that DORB may prove a useful raw material source for the production of anti-obesity peptides which might enhance the therapeutic and commercial performance of functional foods and healthcare products.

Frontiers in Bioengineering and Biotechnology

Graphical A biorefinery based on microalgae promotes a circular-green economic model.

Heliyon

The lingzhi mushroom (Ganoderma lucidum) is well known for its medicinal properties and has long ... more The lingzhi mushroom (Ganoderma lucidum) is well known for its medicinal properties and has long played a role in traditional oriental medicine due to its health-giving benefits and potential to extend life expectancy. The mushroom contains a number of highly bioactive compounds and can also act as an excellent source of protein. This research investigated the peptides obtained from the protein hydrolysates of lingzhi mushrooms to assess their free radical scavenging abilities. These peptides were acquired via different proteases (Alcalase, Neutrase, papain, and pepsin-pancreatin) and were tested at a range of different concentrations (1.0%, 2.5%, and 5.0% w/ v). The highest levels of 2,2 0-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging activities were presented by lingzhi mushroom hydrolysate using 2.5% (w/v) pepsin-pancreatin after 6 h of digestion. The hydrolysate was then fractionated using 10, 5, 3, and 0.65 kDa molecular weight cutoff membranes. The results showed that the MW 0.65 kDa fraction had the highest level of free radical scavenging activity. Further analysis of this MW 0.65 kDa fraction began with another RP-HPLC fractionation technique to obtain three further sub-fractions. De novo peptide sequencing using electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) was chosen as the optimum method for studying the F 3 sub-fraction. DRVSIYGWG and ALLSISSF were discovered as new peptides with different antioxidant properties. Adenocarcinoma colon (Caco-2) cells showed the antioxidant action of these synthesized peptides. This activity was linked to peptide concentration. The peptides and their pure synthetic

Frontiers in Bioengineering and Biotechnology

Over the last several decades, concerns about climate change and pollution due to human activity ... more Over the last several decades, concerns about climate change and pollution due to human activity has gained widespread attention. Microalgae have been proposed as a suitable biological platform to reduce carbon dioxide, a major greenhouse gas, while also creating commercial sources of high-value compounds such as medicines, cosmetics, food, feed, and biofuel. Industrialization of microalgae culture and valorization is still limited by significant challenges in scaling up the production processes due to economic constraints and productivity capacities. Therefore, a boost in resource usage efficiency is required. This enhancement not only lowers manufacturing costs but also enhancing the long-term viability of microalgae-based products. Using wastewater as a nutrient source is a great way to reduce manufacturing costs. Furthermore, water scarcity is one of the most important global challenges. In recent decades, industrialization, globalization, and population growth have all impacted...

High quality RNA products from bacterial cells are required for the molecular study. Sample prepa... more High quality RNA products from bacterial cells are required for the molecular study. Sample preparation to acquire the high-quality RNA especially the Gram-positive bacteria like Bacillus sp., the model organism, remains a critical burden toward the integration of full molecular downstream analyses although several methods have been proposed including conventional or kit-based protocols. Those techniques were simply developed using the cell samples at certain growth stages unless some molecular studies require RNA samples gathered under different physiological stages of growth and process conditions. Herein, we developed the simple yet effective cell-lysis technique prior to RNA extraction by modifying the commercial kit-based protocols. Bacillus subtilis TL7-3 was used as the model organism in this study. Lysozyme loading (20 mg/mL) as well as the incubation time (30 min) and temperature (37 °C) was responsible for cell lysis and increased RNA concentration in the samples. Invert m...

Scientific Reports, 2022

It is anticipated that calcium-chelating peptides may serve to enhance the absorption of calcium.... more It is anticipated that calcium-chelating peptides may serve to enhance the absorption of calcium. This research examined defatted lemon basil seeds (DLBS) which had been treated with Alcalase under optimized parameters for the degree of hydrolysis for proteolysis, discovering that the activity for calcium-binding in a competitive condition with phosphate ion was 60.39 ± 1.545%. The purification of the hydrolysates was performed via ultrafiltration along with reversed-phase high performance liquid chromatography (RP-HPLC). Determination of the purified peptide amino acid sequence was confirmed for both peptides and reported as Ala-Phe-Asn-Arg-Ala-Lys-Ser-Lys-Ala-Leu-Asn-Glu-Asn (AFNRAKSKALNEN; Basil-1), and Tyr-Asp-Ser-Ser-Gly-Gly-Pro-Thr-Pro-Trp-Leu-Ser-Pro-Tyr (YDSSGGPTPWLSPY; Basil-2). The respective activities for calcium-binding were 38.62 ± 1.33%, and 42.19 ± 2.27%. Fluorescence spectroscopy, and fourier transform infrared spectroscopy were employed in order to assess the chela...

RSC Advances, 2020

The isolation and subsequent identification of the two novel ACE-inhibitory peptides, ETSGMKPTEL,... more The isolation and subsequent identification of the two novel ACE-inhibitory peptides, ETSGMKPTEL, and ISSMGILVCL from the longan seeds were achieved. The inhibition mechanism was investigated by molecular docking.

Poultry Science, 2020

This study examined the antioxidant capabilities of peptides derived from chicken feather meal (C... more This study examined the antioxidant capabilities of peptides derived from chicken feather meal (CFM) protein hydrolysates which were produced using 3 different microbial proteases (Neutrase, Alcalase, and flavourzyme) and tested at varying concentrations, namely 1, 2, and 5% by weight. The highest levels of 2,2diphenyl-1-picrylhydrazl (DPPH) and 2,2 0-azino-bis-3ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activities were presented by CFM hydrolysate derived using 5 wt% Neutrase and digested for 4 h. Fractionation of this particular hydrolysate was then performed by applying 10, 5, 3, and 0.65 kDa molecular weight cutoff membranes. It was then determined that the molecular weight (MW) , 0.65 kDa fraction achieved the greatest level of free radical scavenging activity in the context of DPPH and ABTS. The MW , 0.65 kDa fraction then underwent additional fractionation using reverse-phase high-performance liquid chromatography to derive 3 main fractions designated as F 1 , F 2 , and F 3. All of these fractions presented a high level of activity in DPPH radical scavenging, although no significant ABTS scavenging was observed. Quadrupole time-of-flight tandem mass spectrometry was used in determining the peptide contents of the fractions as Phe-Asp-Asp-Arg-Gly-Arg-X for F 1 (FDDRGRX, 875 Da), Val-Thr-Leu-Ala-Val-Thr-Lys-His for F 2 (VTLAVTKH, 868 Da), and Val-Ser-Glu-Ile-X-Ser-Ile-Pro-Ile-Ser for F 3 (VSEIXSI-PIS, 1,055 Da). Moreover, the F 2 fraction was shown to be capable of preventing DNA damage induced by hydroxyl radicals, as indicated in tests using the plasmids pKS, pUC19, and pBR322 via the Fenton reaction. This outcome was demonstrated through in vitro antiproliferative activity in human cell lines based on SW620 colon cancer, using the 3-(4,5-dimethyl-2-thiazolyl)-2,5diphenyl-2H-tetrazolium bromide assay. The F 2 fraction at 0.5 wt.% was also shown to be capable of inducing weak early apoptosis, which could be measured by using the Fluorescein isothiocyanate Annexin V Apoptosis Detection Kit with Propidium Iodide Solution. Furthermore, an increase in caspase-3 and caspase-8 activity was observed in SW620 cells following exposure for 24 h and 48 h.

Food & Function, 2020

Peanut worm (Sipunculus nudus Linn.) protein was hydrolyzed by three proteases and NO scavenging ... more Peanut worm (Sipunculus nudus Linn.) protein was hydrolyzed by three proteases and NO scavenging activity of the protein hydrolysates were evaluated.

Agricultural Water Management, 2019

Nitrogen-rich wastewater is a major problem for the aquaculture industry. To investigate whether ... more Nitrogen-rich wastewater is a major problem for the aquaculture industry. To investigate whether algae could be used to remove ammonium from brackish shrimp aquaculture wastewater, marine cyanobacterium Synechococcus sp. VDW was cultured in BG11 medium supplemented with Turks Island salt solution and different concentrations of NH 4 Cl (1.0-40.0 mg L −1) for 18 days. The cell density of the Synechococcus sp. VDW cultures increased in medium containing between 1 and 10 mg L −1 of NH 4 Cl, while ammonium concentrations greater than 20.0 mg L −1 had a negative effect on growth. Glutamine synthetase and glutamate synthase activities were also examined, and were found to increase with cell density. Meanwhile, glutamate dehydrogenase activity increased in response to high NH 4 Cl concentrations (20.0-40.0 mg L −1). The cellular response to ammonium excess was confirmed by measuring gene expression levels using quantitative PCR. Expression of both glnA and gltB was down-regulated compared with the control, while that of gdhA was up-regulated. At an initial concentration of 1-10 mg L −1 , 98-100% of the ammonium was removed by day 6 of cultivation. Therefore, these findings suggest that Synechococcus sp. VDW can remove ammonium from contaminated brackish water, and may be helpful for improving the quality of aquaculture wastewater.

Journal of Oleo Science, 2019

grow without using arable land. Moreover, algae can effectively capture carbon dioxide through ph... more grow without using arable land. Moreover, algae can effectively capture carbon dioxide through photosynthesis and produce polysaccharides and triacylglycerol TAG. These compounds are the raw materials for production of bioethanol and biodiesel, which can be used in current engines without major modifications 2, 3. Although, many algae are accumulate significant quantities of TAG 20-50 of total dry weight and has potential as a raw material for biodiesel production than cyanobacteria, which typically store carbon as glycogen and or polyhydroxybutyrate PHB .

Journal of Evolutionary Biochemistry and Physiology, 2020

Abstract Aspergillus terreus , an acid tolerant fungus that grows over a wide range of pH from 2 ... more Abstract Aspergillus terreus , an acid tolerant fungus that grows over a wide range of pH from 2 to 8, was employed in this study. Submerged fermentation was conducted under low dissolved oxygen (10% DO) and pH (pH 2). The fermentation kinetics was investigated together with transcriptome to identify key genes associated with metabolic responses to such environmental stresses by high-throughput RNA-seq technology. It was clear that under low dissolved oxygen, glycolysis was uncoupled from oxidative phosphorylation due to limited ATP regeneration, as observed from the downregulation of genes in the mitochondrial respiratory chain complex and slow growth. On the contrary, the transcriptional levels of genes encoding ATPases and cation transporters remained unchanged at –1 ≤ log2 fold change ≤ 1, revealing that A. terreus possessed an excellent defense mechanism to low pH. In addition, genes associated with cell wall synthesis and repair, including chitin synthase and 1,3-beta-glucan synthase genes, and genes in the cluster for cell wall protection and repair were upregulated. This evidence clearly demonstrated that A. terreus was able to survive under the acidic environment. In summary, we comprehensively analyzed and discussed the transcriptome of A. terreus . The findings obtained here provide a theoretical basis for better understaning the molecular mechanism of the fungal response to the environmental factors, providing thereby an insight into the process of designing and optimizing the production of biological products of interest. This approach can be applicable to other metabolic pathways of interest in particular microorganisms.

When fish are processed, fish bone becomes a key component of the waste, but to date very few res... more When fish are processed, fish bone becomes a key component of the waste, but to date very few researchers have sought to use fish bone to prepare protein hydrolysates as a means of adding value to the final product. This study, therefore, examines the potential of salmon bone, through an analysis of the benefits of its constituent components, namely fat, moisture, protein, and ash. In particular, the study seeks to optimize the process of enzymatic hydrolysis of salmon bone with trypsin in order to produce angiotensin-I converting enzyme (ACE) inhibitory peptides making use of response surface methodology in combination with central composite design (CCD). Optimum hydrolysis conditions concerning DH (degree of hydrolysis) and ACE-inhibitory activity were initially determined using the response surface model. Having thus determined which of the salmon bone protein hydrolysates (SBPH) offered the greatest level of ACE-inhibitory activity, these SBPH were duly selected to undergo ultra...

RSC Advances

De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry.

Molecular Biology Reports

BACKGROUND High quality RNA is required for the molecular study. Sample preparation of the spore-... more BACKGROUND High quality RNA is required for the molecular study. Sample preparation of the spore-forming, Gram-positive bacteria like Bacillus sp., remains challenging although several methods have been proposed. Those techniques were simply developed using cell samples at certain growth stages despite some molecular studies like transcriptomic analyses require RNA samples from different physiological stages. METHODS AND RESULTS We developed the rapid, simple yet effective cell-lysis technique with limit use of harsh reagents by modifying the kit-based protocols. Appropriate lysozyme loading (20 mg/mL), incubation time (30 min), and temperature (37 °C) enabled cell lysis and enhanced RNA extraction from both vegetative cells and endospores of Bacillus subtilis TL7-3. High RNA Integrity Numbers and ratios of A260/A280 and A260/A230 of all RNA products collected during the batch cultivation confirmed that invert mixing with absolute ethanol prevented RNA damage during protein denaturation. With the process modification of the major steps in cell lysis and RNA extraction compared with the kit-based protocols that are typically used in laboratory work, interestingly, our modified protocol, simple-yet-effective, yielded higher concentration, purity, and integrity of RNA products from all cell samples collected at different physiological stages. While the kit-based protocols either failed to provide high RNA concentration or RNA purity and integrity for all cell samples particularly during the late-log, stationary, or sporulation. CONCLUSIONS Therefore, we can claim the significance of this modified protocol to be applicable for RNA extraction to those spore-forming Gram-positive bacteria not limited to B. subtilis growing at varied physiological stages.

Food & Function

This research examined the antioxidant abilities of peptides derived from the Hericium erinaceus ... more This research examined the antioxidant abilities of peptides derived from the Hericium erinaceus mushroom produced via three microbial proteases at varying concentrations.

Food & Function

The study determines optimized process conditions to maximize ACE inhibitory peptide production. ... more The study determines optimized process conditions to maximize ACE inhibitory peptide production. The two novel hexa-peptides (LGRNLPPI and GPAGPAGL) from de-fatted lemon basil seeds (DLBS) was achieved.

PLOS ONE, Sep 2, 2021

When fish are processed, fish bone becomes a key component of the waste, but to date very few res... more When fish are processed, fish bone becomes a key component of the waste, but to date very few researchers have sought to use fish bone to prepare protein hydrolysates as a means of adding value to the final product. This study, therefore, examines the potential of salmon bone, through an analysis of the benefits of its constituent components, namely fat, moisture, protein, and ash. In particular, the study seeks to optimize the process of enzymatic hydrolysis of salmon bone with trypsin in order to produce angiotensin-I converting enzyme (ACE) inhibitory peptides making use of response surface methodology in combination with central composite design (CCD). Optimum hydrolysis conditions concerning DH (degree of hydrolysis) and ACE-inhibitory activity were initially determined using the response surface model. Having thus determined which of the salmon bone protein hydrolysates (SBPH) offered the greatest level of ACE-inhibitory activity, these SBPH were duly selected to undergo ultrafiltration for further fractionation. It was found that the greatest ACE-inhibitory activity was achieved by the SBPH fraction which had a molecular weight lower than 0.65 kDa. This fraction underwent further purification using RP-HPLC, revealing that the F 7 fraction offered the best ACE-inhibitory activity. For ACE inhibition, the ideal peptide in the context of the F 7 fraction comprised eight amino acids: Phe-Cys-Leu-Tyr-Glu-Leu-Ala-Arg (FCLYELAR), while analysis of the Lineweaver-Burk plot revealed that the FCLYELAR peptide can serve as an uncompetitive ACE inhibitor. An examination of the molecular docking process showed that the FCLYELAR peptide was primarily able to provide ACE-inhibitory qualities as a consequence of the hydrogen bond interactions taking place between ACE and the peptide. Furthermore, upon isolation form the SBPH, the ACEinhibitory peptide demonstrated ACE-inhibitory capabilities in vitro, underlining its potential for applications in the food and pharmaceutical sectors.

RSC Advances, 2021

De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry. DORB ... more De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry. DORB may prove to be an important protein source, and also contains many other micronutrients. This study has the principal aim of optimizing the process of DORB protein hydrolysate preparation, and then testing the hydrolysate to determine its lipase inhibitory activity. DORB underwent hydrolysis using Alcalase® and response surface methodology (RSM). The resulting degree of hydrolysis (DH) was then monitored along with the extent of any lipase inhibitory activity. The optimum levels of lipase inhibition were obtained at a temperature of 49.88 C, a duration of 150.43 minutes, and 1.53% Alcalase® used for the sample 5% (w/v) solution. In these conditions, the DH value was 35.65%, and the IC 50 value for lipase inhibitory activity was 2.84 mg mL À1. Five ranges of different molecular weights were obtained via fractionation, whereupon it was determined that the highest level of inhibitory activity was achieved by the <0.65 kDa fraction. This fraction was then further purified via RP-HPLC, and the resulting peak had a retention time of 21.75 minutes (F 2 sub-fraction) and exhibited high lipase inhibitory activity. Mass spectrometry was used to determine the amino acid sequence for this peak, identified as FYLGYCDY. This particular peptide is categorized as bitter, with a non-toxic profile, and having poor water solubility. The synthesized form of this peptide showed lipase inhibitory activity measured by an IC 50 value of 0.47 AE 0.02 mM. The Lineweaver-Burk plot revealed that FYLGYCDY is a non-competitive inhibitor, while analysis of the docking results provided details of the FYLGYCDY peptide binding site with the porcine pancreatic lipase (PPL) complex, which is a competitive type. It can be inferred from these findings that DORB may prove a useful raw material source for the production of anti-obesity peptides which might enhance the therapeutic and commercial performance of functional foods and healthcare products.

Frontiers in Bioengineering and Biotechnology

Graphical A biorefinery based on microalgae promotes a circular-green economic model.

Heliyon

The lingzhi mushroom (Ganoderma lucidum) is well known for its medicinal properties and has long ... more The lingzhi mushroom (Ganoderma lucidum) is well known for its medicinal properties and has long played a role in traditional oriental medicine due to its health-giving benefits and potential to extend life expectancy. The mushroom contains a number of highly bioactive compounds and can also act as an excellent source of protein. This research investigated the peptides obtained from the protein hydrolysates of lingzhi mushrooms to assess their free radical scavenging abilities. These peptides were acquired via different proteases (Alcalase, Neutrase, papain, and pepsin-pancreatin) and were tested at a range of different concentrations (1.0%, 2.5%, and 5.0% w/ v). The highest levels of 2,2 0-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging activities were presented by lingzhi mushroom hydrolysate using 2.5% (w/v) pepsin-pancreatin after 6 h of digestion. The hydrolysate was then fractionated using 10, 5, 3, and 0.65 kDa molecular weight cutoff membranes. The results showed that the MW 0.65 kDa fraction had the highest level of free radical scavenging activity. Further analysis of this MW 0.65 kDa fraction began with another RP-HPLC fractionation technique to obtain three further sub-fractions. De novo peptide sequencing using electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS/MS) was chosen as the optimum method for studying the F 3 sub-fraction. DRVSIYGWG and ALLSISSF were discovered as new peptides with different antioxidant properties. Adenocarcinoma colon (Caco-2) cells showed the antioxidant action of these synthesized peptides. This activity was linked to peptide concentration. The peptides and their pure synthetic

Frontiers in Bioengineering and Biotechnology

Over the last several decades, concerns about climate change and pollution due to human activity ... more Over the last several decades, concerns about climate change and pollution due to human activity has gained widespread attention. Microalgae have been proposed as a suitable biological platform to reduce carbon dioxide, a major greenhouse gas, while also creating commercial sources of high-value compounds such as medicines, cosmetics, food, feed, and biofuel. Industrialization of microalgae culture and valorization is still limited by significant challenges in scaling up the production processes due to economic constraints and productivity capacities. Therefore, a boost in resource usage efficiency is required. This enhancement not only lowers manufacturing costs but also enhancing the long-term viability of microalgae-based products. Using wastewater as a nutrient source is a great way to reduce manufacturing costs. Furthermore, water scarcity is one of the most important global challenges. In recent decades, industrialization, globalization, and population growth have all impacted...

High quality RNA products from bacterial cells are required for the molecular study. Sample prepa... more High quality RNA products from bacterial cells are required for the molecular study. Sample preparation to acquire the high-quality RNA especially the Gram-positive bacteria like Bacillus sp., the model organism, remains a critical burden toward the integration of full molecular downstream analyses although several methods have been proposed including conventional or kit-based protocols. Those techniques were simply developed using the cell samples at certain growth stages unless some molecular studies require RNA samples gathered under different physiological stages of growth and process conditions. Herein, we developed the simple yet effective cell-lysis technique prior to RNA extraction by modifying the commercial kit-based protocols. Bacillus subtilis TL7-3 was used as the model organism in this study. Lysozyme loading (20 mg/mL) as well as the incubation time (30 min) and temperature (37 °C) was responsible for cell lysis and increased RNA concentration in the samples. Invert m...

Scientific Reports, 2022

It is anticipated that calcium-chelating peptides may serve to enhance the absorption of calcium.... more It is anticipated that calcium-chelating peptides may serve to enhance the absorption of calcium. This research examined defatted lemon basil seeds (DLBS) which had been treated with Alcalase under optimized parameters for the degree of hydrolysis for proteolysis, discovering that the activity for calcium-binding in a competitive condition with phosphate ion was 60.39 ± 1.545%. The purification of the hydrolysates was performed via ultrafiltration along with reversed-phase high performance liquid chromatography (RP-HPLC). Determination of the purified peptide amino acid sequence was confirmed for both peptides and reported as Ala-Phe-Asn-Arg-Ala-Lys-Ser-Lys-Ala-Leu-Asn-Glu-Asn (AFNRAKSKALNEN; Basil-1), and Tyr-Asp-Ser-Ser-Gly-Gly-Pro-Thr-Pro-Trp-Leu-Ser-Pro-Tyr (YDSSGGPTPWLSPY; Basil-2). The respective activities for calcium-binding were 38.62 ± 1.33%, and 42.19 ± 2.27%. Fluorescence spectroscopy, and fourier transform infrared spectroscopy were employed in order to assess the chela...

RSC Advances, 2020

The isolation and subsequent identification of the two novel ACE-inhibitory peptides, ETSGMKPTEL,... more The isolation and subsequent identification of the two novel ACE-inhibitory peptides, ETSGMKPTEL, and ISSMGILVCL from the longan seeds were achieved. The inhibition mechanism was investigated by molecular docking.

Poultry Science, 2020

This study examined the antioxidant capabilities of peptides derived from chicken feather meal (C... more This study examined the antioxidant capabilities of peptides derived from chicken feather meal (CFM) protein hydrolysates which were produced using 3 different microbial proteases (Neutrase, Alcalase, and flavourzyme) and tested at varying concentrations, namely 1, 2, and 5% by weight. The highest levels of 2,2diphenyl-1-picrylhydrazl (DPPH) and 2,2 0-azino-bis-3ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activities were presented by CFM hydrolysate derived using 5 wt% Neutrase and digested for 4 h. Fractionation of this particular hydrolysate was then performed by applying 10, 5, 3, and 0.65 kDa molecular weight cutoff membranes. It was then determined that the molecular weight (MW) , 0.65 kDa fraction achieved the greatest level of free radical scavenging activity in the context of DPPH and ABTS. The MW , 0.65 kDa fraction then underwent additional fractionation using reverse-phase high-performance liquid chromatography to derive 3 main fractions designated as F 1 , F 2 , and F 3. All of these fractions presented a high level of activity in DPPH radical scavenging, although no significant ABTS scavenging was observed. Quadrupole time-of-flight tandem mass spectrometry was used in determining the peptide contents of the fractions as Phe-Asp-Asp-Arg-Gly-Arg-X for F 1 (FDDRGRX, 875 Da), Val-Thr-Leu-Ala-Val-Thr-Lys-His for F 2 (VTLAVTKH, 868 Da), and Val-Ser-Glu-Ile-X-Ser-Ile-Pro-Ile-Ser for F 3 (VSEIXSI-PIS, 1,055 Da). Moreover, the F 2 fraction was shown to be capable of preventing DNA damage induced by hydroxyl radicals, as indicated in tests using the plasmids pKS, pUC19, and pBR322 via the Fenton reaction. This outcome was demonstrated through in vitro antiproliferative activity in human cell lines based on SW620 colon cancer, using the 3-(4,5-dimethyl-2-thiazolyl)-2,5diphenyl-2H-tetrazolium bromide assay. The F 2 fraction at 0.5 wt.% was also shown to be capable of inducing weak early apoptosis, which could be measured by using the Fluorescein isothiocyanate Annexin V Apoptosis Detection Kit with Propidium Iodide Solution. Furthermore, an increase in caspase-3 and caspase-8 activity was observed in SW620 cells following exposure for 24 h and 48 h.

Food & Function, 2020

Peanut worm (Sipunculus nudus Linn.) protein was hydrolyzed by three proteases and NO scavenging ... more Peanut worm (Sipunculus nudus Linn.) protein was hydrolyzed by three proteases and NO scavenging activity of the protein hydrolysates were evaluated.

Agricultural Water Management, 2019

Nitrogen-rich wastewater is a major problem for the aquaculture industry. To investigate whether ... more Nitrogen-rich wastewater is a major problem for the aquaculture industry. To investigate whether algae could be used to remove ammonium from brackish shrimp aquaculture wastewater, marine cyanobacterium Synechococcus sp. VDW was cultured in BG11 medium supplemented with Turks Island salt solution and different concentrations of NH 4 Cl (1.0-40.0 mg L −1) for 18 days. The cell density of the Synechococcus sp. VDW cultures increased in medium containing between 1 and 10 mg L −1 of NH 4 Cl, while ammonium concentrations greater than 20.0 mg L −1 had a negative effect on growth. Glutamine synthetase and glutamate synthase activities were also examined, and were found to increase with cell density. Meanwhile, glutamate dehydrogenase activity increased in response to high NH 4 Cl concentrations (20.0-40.0 mg L −1). The cellular response to ammonium excess was confirmed by measuring gene expression levels using quantitative PCR. Expression of both glnA and gltB was down-regulated compared with the control, while that of gdhA was up-regulated. At an initial concentration of 1-10 mg L −1 , 98-100% of the ammonium was removed by day 6 of cultivation. Therefore, these findings suggest that Synechococcus sp. VDW can remove ammonium from contaminated brackish water, and may be helpful for improving the quality of aquaculture wastewater.

Journal of Oleo Science, 2019

grow without using arable land. Moreover, algae can effectively capture carbon dioxide through ph... more grow without using arable land. Moreover, algae can effectively capture carbon dioxide through photosynthesis and produce polysaccharides and triacylglycerol TAG. These compounds are the raw materials for production of bioethanol and biodiesel, which can be used in current engines without major modifications 2, 3. Although, many algae are accumulate significant quantities of TAG 20-50 of total dry weight and has potential as a raw material for biodiesel production than cyanobacteria, which typically store carbon as glycogen and or polyhydroxybutyrate PHB .

Journal of Evolutionary Biochemistry and Physiology, 2020

Abstract Aspergillus terreus , an acid tolerant fungus that grows over a wide range of pH from 2 ... more Abstract Aspergillus terreus , an acid tolerant fungus that grows over a wide range of pH from 2 to 8, was employed in this study. Submerged fermentation was conducted under low dissolved oxygen (10% DO) and pH (pH 2). The fermentation kinetics was investigated together with transcriptome to identify key genes associated with metabolic responses to such environmental stresses by high-throughput RNA-seq technology. It was clear that under low dissolved oxygen, glycolysis was uncoupled from oxidative phosphorylation due to limited ATP regeneration, as observed from the downregulation of genes in the mitochondrial respiratory chain complex and slow growth. On the contrary, the transcriptional levels of genes encoding ATPases and cation transporters remained unchanged at –1 ≤ log2 fold change ≤ 1, revealing that A. terreus possessed an excellent defense mechanism to low pH. In addition, genes associated with cell wall synthesis and repair, including chitin synthase and 1,3-beta-glucan synthase genes, and genes in the cluster for cell wall protection and repair were upregulated. This evidence clearly demonstrated that A. terreus was able to survive under the acidic environment. In summary, we comprehensively analyzed and discussed the transcriptome of A. terreus . The findings obtained here provide a theoretical basis for better understaning the molecular mechanism of the fungal response to the environmental factors, providing thereby an insight into the process of designing and optimizing the production of biological products of interest. This approach can be applicable to other metabolic pathways of interest in particular microorganisms.

When fish are processed, fish bone becomes a key component of the waste, but to date very few res... more When fish are processed, fish bone becomes a key component of the waste, but to date very few researchers have sought to use fish bone to prepare protein hydrolysates as a means of adding value to the final product. This study, therefore, examines the potential of salmon bone, through an analysis of the benefits of its constituent components, namely fat, moisture, protein, and ash. In particular, the study seeks to optimize the process of enzymatic hydrolysis of salmon bone with trypsin in order to produce angiotensin-I converting enzyme (ACE) inhibitory peptides making use of response surface methodology in combination with central composite design (CCD). Optimum hydrolysis conditions concerning DH (degree of hydrolysis) and ACE-inhibitory activity were initially determined using the response surface model. Having thus determined which of the salmon bone protein hydrolysates (SBPH) offered the greatest level of ACE-inhibitory activity, these SBPH were duly selected to undergo ultra...

RSC Advances

De-oiled rice bran (DORB) is a potentially useful by-product of the rice bran oil industry.

Molecular Biology Reports

BACKGROUND High quality RNA is required for the molecular study. Sample preparation of the spore-... more BACKGROUND High quality RNA is required for the molecular study. Sample preparation of the spore-forming, Gram-positive bacteria like Bacillus sp., remains challenging although several methods have been proposed. Those techniques were simply developed using cell samples at certain growth stages despite some molecular studies like transcriptomic analyses require RNA samples from different physiological stages. METHODS AND RESULTS We developed the rapid, simple yet effective cell-lysis technique with limit use of harsh reagents by modifying the kit-based protocols. Appropriate lysozyme loading (20 mg/mL), incubation time (30 min), and temperature (37 °C) enabled cell lysis and enhanced RNA extraction from both vegetative cells and endospores of Bacillus subtilis TL7-3. High RNA Integrity Numbers and ratios of A260/A280 and A260/A230 of all RNA products collected during the batch cultivation confirmed that invert mixing with absolute ethanol prevented RNA damage during protein denaturation. With the process modification of the major steps in cell lysis and RNA extraction compared with the kit-based protocols that are typically used in laboratory work, interestingly, our modified protocol, simple-yet-effective, yielded higher concentration, purity, and integrity of RNA products from all cell samples collected at different physiological stages. While the kit-based protocols either failed to provide high RNA concentration or RNA purity and integrity for all cell samples particularly during the late-log, stationary, or sporulation. CONCLUSIONS Therefore, we can claim the significance of this modified protocol to be applicable for RNA extraction to those spore-forming Gram-positive bacteria not limited to B. subtilis growing at varied physiological stages.

Food & Function

This research examined the antioxidant abilities of peptides derived from the Hericium erinaceus ... more This research examined the antioxidant abilities of peptides derived from the Hericium erinaceus mushroom produced via three microbial proteases at varying concentrations.

Food & Function

The study determines optimized process conditions to maximize ACE inhibitory peptide production. ... more The study determines optimized process conditions to maximize ACE inhibitory peptide production. The two novel hexa-peptides (LGRNLPPI and GPAGPAGL) from de-fatted lemon basil seeds (DLBS) was achieved.