Kinetics and Modeling of Pepsin Soluble Collagen (PSC) Extraction from the Skin of Malaysian catfish (Hybrid Clarias sp.)

Peck Loo Kiew,Mat Don Mashitah,Zainal Ahmad 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.1

The empirical kinetic models for the pepsin solublecollagen extraction from the skin of cultured hybrid catfish (Clarissp.) were studied using four two-parametric models, namely thepower law, parabolic diffusion, Peleg’s and Elovich’s models. ThePeleg’s model was found to be the best model capable of predictingthe extraction data with R2 >0.9, p-value >2.0%, and RMSD<10.0%, respectively. Kinetic models based on the second orderrate equation were successfully developed to describe the extractionprocesses with different processing variables. Extraction rateconstant, initial extraction rate and equilibrium concentrations fordifferent acetic acid concentrations, liquid to solid ratios, andpepsin dosages were predicted. The verification of the developedmodels showed that the experimental values agreed with thepredicted ones, with percentage error differences in the range of0.03–3.91%.

Malaysian Freshwater Fish: Exploitable Source for Collagen Extraction

Peck Loo Kiew,Mashitah Mat Don 중소기업융합학회 2015 중소기업융합학회 국제학술대회논문집 Vol.2 No.1

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were extracted from the skin of cultured hybrid catfish (Clarias gariepinus X Clarias macrocephalus), red tilapia (Oreochromis niloticus), black tilapia (Oreochromis mossambicus), pangasius catfish (Pangasianodon hypophthalmus), sultan fish (Leptobarbus hoevenii), and labyrinth fish (Trichogaster trichopterus), freshwater fishes that are widely consumed in Malaysia. The extracted yields for all selected species were higher for PSC as compared to ASC. Among all, catfish of the hybrid Clarias sp. contained the highest amount of skin collagen, both ASC and PSC at 36.73 ± 0.51 % and 58.17 ± 0.54 %, respectively. The extracted collagens were confirmed as type I collagen based on the SDSPAGE analysis and amino acid composition. Both ASC and PSC had glycine as the major amino acid and no cysteine was found in any of the extracted collagen. This was consistent with reported characteristics of type I collagen in the literature. Presence of collagen in the skin of these fishes showed the possibility to convert these natural resources into value-added product such as collagen, thus aids in improving their commercial values whilst assisting in ensuring the sustainability of Malaysian freshwater fishes.

Kinetics and Modeling of Pepsin Soluble Collagen (PSC) Extraction from the Skin of Malaysian catfish (Hybrid Clarias sp.)

Kiew, Peck Loo,Mashitah, Mat Don,Ahmad, Zainal The Korean Society for Applied Biological Chemistr 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.1

The empirical kinetic models for the pepsin soluble collagen extraction from the skin of cultured hybrid catfish (Claris sp.) were studied using four two-parametric models, namely the power law, parabolic diffusion, Peleg's and Elovich's models. The Peleg's model was found to be the best model capable of predicting the extraction data with $R^2$ >0.9, p-value >2.0%, and RMSD <10.0%, respectively. Kinetic models based on the second order rate equation were successfully developed to describe the extraction processes with different processing variables. Extraction rate constant, initial extraction rate and equilibrium concentrations for different acetic acid concentrations, liquid to solid ratios, and pepsin dosages were predicted. The verification of the developed models showed that the experimental values agreed with the predicted ones, with percentage error differences in the range of 0.03-3.91%.

Optimization of Process Variables for the Synthesis of Silver Nanoparticles by Pycnoporus sanguineus using Statistical Experimental Design

Yen San Chan,Mashitah Mat Don 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.1

Sequential optimization strategy based on statistical experimental design and one-factor-at-a-time (OFAT) method were employed to optimize the process parameters for the enhancement of silver nanoparticles (AgNPs) production through biological synthesis using Pycnoporus sanguineus. Based on the OFAT method, three significant components influencing the size of AgNPs produced were identified as AgNO3 concentration,incubation temperature, and agitation speed. The optimum values of these process parameter for the synthesis of AgNPs were determined using response surface methodology (RSM) based on Box-Behnken design. The validity of the model developed was verified, and the statistical analysis showed that the optimum operating conditions were 0.001 M of AgNO3, 38oC, and 200 rpm with the smallest AgNPs produced at 14.86 nm. The disc diffusion method also suggested that AgNPs produced using optimum conditions have higher antimicrobial activity compared to the unoptimized AgNPs. The present study developed a robust operating condition for the production of AgNPs by P. sanguineus, which was 8.6-fold smaller than that obtained from un-optimized conditions.

Flavonoid Production by T. lactinea: Screening of Culture Conditions Via OFAT and Optimization Using Response Surface Methodology (RSM)

Yus Azila Yahaya,Mashitah Mat Don 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

The culture conditions for flavonoid production by thefungus Trametes lactinea was screened using one-factor-at-a-time(OFAT) and optimized with response surface methodology. T. Lactinea was found to grow well in Medium 3, and the highestflavonoid was produced in Medium 2. Medium 2 contains yeastextract, glucose, KH2PO4, Na2HPO4 · 12H2O, MgSO4 · 7H2O, and(NH4)2 · SO4, whereas Medium 3 contains yeast extract, glucoseand peptone. Four selected parameters in OFAT were furtheroptimized for flavonoid production by T. lactinea using responsesurface methodology via Box Behnken design (BBD). Interactionsof culture conditions and optimization of the system were thenstudied using BBD with four levels of the four variables in a batchflask culture. Experimentation showed that the model developedbased on BBD had predicted flavonoid production with R2=0.9983. The predicted flavonoid production was optimum (20.02μg/mL) when the culture conditions were at 8.1 days of incubationperiod, 5.2 mL of inoculum, incubation temperature at 34.6oC,and the production medium with initial pH 6. The results indicatedthat BBD method was effective in optimizing the culture conditionsof flavonoid production by T.lactinea.

Flavonoid Production by T. lactinea: Screening of Culture Conditions Via OFAT and Optimization Using Response Surface Methodology (RSM)

Yahaya, Yus Azila,Don, Mashitah Mat 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

The culture conditions for flavonoid production by the fungus Trametes lactinea was screened using one-factor-at-a-time (OFAT) and optimized with response surface methodology. T. Lactinea was found to grow well in Medium 3, and the highest flavonoid was produced in Medium 2. Medium 2 contains yeast extract, glucose, $KH_2PO_4$, $Na_2HPO_4{\cdot}12H_2O$, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2{\cdot}SO_4$, whereas Medium 3 contains yeast extract, glucose and peptone. Four selected parameters in OFAT were further optimized for flavonoid production by T. lactinea using response surface methodology via Box Behnken design (BBD). Interactions of culture conditions and optimization of the system were then studied using BBD with four levels of the four variables in a batch flask culture. Experimentation showed that the model developed based on BBD had predicted flavonoid production with $R^2=0.9983$. The predicted flavonoid production was optimum ($20.02{\mu}g/mL$) when the culture conditions were at 8.1 days of incubation period, 5.2 mL of inoculum, incubation temperature at $34.6^{\circ}C$, and the production medium with initial pH 6. The results indicated that BBD method was effective in optimizing the culture conditions of flavonoid production by T.lactinea.

Isolation and Characterization of Collagen from the Skin of Malaysian catfish (Hybrid Clarias sp.)

Peck Loo Kiew,Mat Don Mashitah 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.4

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from the skin of hybrid Clarias sp. with the yields of 18.11±0.32 and 26.69±0.54% (wet weight basis), respectively. Both collagens were characterized as type I collagen, containing α1 and α2 chains. Presence of high molecular weight crosslinks were observed in the gel electrophoresis of both collagens. Fourier transform infrared spectra of both collagens were almost similar, suggesting that pepsin hydrolysis did not disrupt the triple helical structure. The amino acid analysis showed glycine was the most abundant, with 207/1000 and 223/1000 residues present in ASC and PSC, respectively. The amounts of imino acids were 185/1000 residues for both. Thermal denaturation temperatures were determined to be 31.5 and 31.0oC, respectively. Both collagens exhibited high solubility in acidic pH (1–5) and below 4% (w/v) NaCl concentration.

Isolation and Characterization of Collagen from the Skin of Malaysian catfish (Hybrid Clarias sp.)

Kiew, Peck Loo,Mashitah, Mat Don 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.4

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from the skin of hybrid Clarias sp. with the yields of $18.11{\pm}0.32$ and $26.69{\pm}0.54%$ (wet weight basis), respectively. Both collagens were characterized as type I collagen, containing ${\alpha}1$ and ${\alpha}2$ chains. Presence of high molecular weight crosslinks were observed in the gel electrophoresis of both collagens. Fourier transform infrared spectra of both collagens were almost similar, suggesting that pepsin hydrolysis did not disrupt the triple helical structure. The amino acid analysis showed glycine was the most abundant, with 207/1000 and 223/1000 residues present in ASC and PSC, respectively. The amounts of imino acids were 185/1000 residues for both. Thermal denaturation temperatures were determined to be 31.5 and $31.0^{\circ}C$, respectively. Both collagens exhibited high solubility in acidic pH (1-5) and below 4% (w/v) NaCl concentration.

Optimization of Process Variables for the Synthesis of Silver Nanoparticles by Pycnoporus sanguineus using Statistical Experimental Design

Chan, Yen San,Don, Mashitah Mat 한국응용생명화학회 2013 Applied Biological Chemistry (Appl Biol Chem) Vol.56 No.1

Sequential optimization strategy based on statistical experimental design and one-factor-at-a-time (OFAT) method were employed to optimize the process parameters for the enhancement of silver nanoparticles (AgNPs) production through biological synthesis using Pycnoporus sanguineus. Based on the OFAT method, three significant components influencing the size of AgNPs produced were identified as $AgNO_3$ concentration, incubation temperature, and agitation speed. The optimum values of these process parameter for the synthesis of AgNPs were determined using response surface methodology (RSM) based on Box-Behnken design. The validity of the model developed was verified, and the statistical analysis showed that the optimum operating conditions were 0.001 M of $AgNO_3$, $38^{\circ}C$, and 200 rpm with the smallest AgNPs produced at 14.86 nm. The disc diffusion method also suggested that AgNPs produced using optimum conditions have higher antimicrobial activity compared to the unoptimized AgNPs. The present study developed a robust operating condition for the production of AgNPs by P. sanguineus, which was 8.6-fold smaller than that obtained from un-optimized conditions.

A Hybrid of Back Propagation Neural Network and Genetic Algorithm for Optimization of Collagen Extraction from Malaysian Cultured Catfish (Hybrid Clarias sp.)

Peck Loo Kiew,Zainal Ahmad,Mashitah Mat Don 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2

Fractional factorial design (FFD) was applied to evaluate the effects of various process parameters in influencing the extraction efficiency of pepsin soluble collagen (PSC) from muscles of cultured catfish (Clarias gariepinus×C. macrocephalus). Result of the first order factorial design showed that acetic acid concentration, acid extraction time, acetic acid to muscles ratio, and stirring speed posed significant effect (P<0.05) on the yield of PSC obtained at the end of the extraction process. Two different artificial intelligence techniques namely artificial neural network (ANN) and genetic algorithm (GA) were then integrated for optimizing the extraction conditions to obtain the highest yield of PSC. The ANN was trained using the back propagation algorithm. A model was successfully generated with R2 value of 0.9527 and MSE value of 0.1672 for unseen data set, implying a good generalization of the network. Input parameters of the established ANN model were subsequently optimized using GA. The hybrid of ANN-GA model predicted a maximum extraction yield of PSC at 238.25 mg/g under the following conditions: an acetic acid concentration of 0.70 M, the acetic acid to muscles ratio of 25.78 mL/g,and the stirring speed of 432.50 rpm. Verification of the optimization showed the percentage error differences between the experimental and predicted values were less than 5%, indicating excellent modeling, predicting ability and optimization by the ANN-GA model.