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Partition efficiency of cytochrome c with alcohol/salt aqueous biphasic flotation system

Ng HS, Ng TC, Kee PE, Tan JS, Yim HS, Lan JC

J Biosci Bioeng, 2020 Feb;129(2):237-241.
PMID: 31629635 DOI: 10.1016/j.jbiosc.2019.08.013

Aqueous biphasic flotation (ABF) integrates aqueous biphasic system (ABS) and solvent sublation for recovery of target biomolecules. The feasibility of the alcohol/salt ABF for exclusive partition of cytochrome c to one specific phase of the system was investigated. Aliphatic alcohols of different carbon chain length (ethanol, 1-propanol and 2-propanol) and salts (sulfate, phosphate and citrate) were used for the phase formation. The effects of phase composition, concentration of sample loading, pH, flotation time and flow rate of the system on the partition efficiency of cytochrome c were determined. Cytochrome c was exclusively partitioned to the alcohol-rich top phase of the ABF of 18% (w/w) ethanol and 26% (w/w) ammonium sulfate with pH 6 and 20% (w/w) of sample loading. Highest partition coefficient (K) of 6.85 ± 0.21 and yield (YT) of 99.40% ± 0.02 were obtained with optimum flotation rate of 10 mL/min and flow rate of 10 min.

Matched MeSH terms: 1-Propanol/chemistry
Fulltext The employment of q-PCR using specific primer targeting on mitochondrial cytochrome-b gene for identification of wild boar meat in meatball samples

Aina GQ, Erwanto Y, Hossain M, Johan MR, Ali ME, Rohman A

J Adv Vet Anim Res, 2019 Sep;6(3):300-307.
PMID: 31583226 DOI: 10.5455/javar.2019.f348

Objective: The objective of this study was to employ real-time or quantitative polymerase chain reaction (q-PCR) using novel species specific primer (SSP) targeting on mitochondrial cytochrome-b of wild boar species (CYTBWB2-wb) gene for the identification of non-halal meat of wild boar meat (WBM) in meatball products.
Materials and Methods: The novel SSP of CYTBWB2-wb was designed by our group using PRIMERQUEST and NCBI software. DNA was extracted using propanol-chloroform-isoamyl alcohol method. The designed SSP was further subjected for validation protocols using DNA isolated from fresh meat and from meatball, which include specificity test, determination of efficiency, limit of detection and repeatability, and application of developed method for analysis of commercially meatball samples.
Results: The results showed that CYTBWB2-wb was specific to wild boar species against other animal species with optimized annealing temperature of 59°C. The efficiency of q-PCR obtained was 91.9% which is acceptable according to the Codex Allimentarius Commission (2010). DNA, with as low as 5 pg/μl, could be detected using q-PCR with primer of CYTBWB2-wb. The developed method was also used for DNA analysis extracted from meatball samples commercially available.
Conclusion: q-PCR using CYTBWB2-wb primers targeting on mitochondrial cytochrome-b gene (forward: CGG TTC CCT CTT AGG CAT TT; Reverse: GGA TGA ACA GGC AGA TGA AGA) can be fruitfully used for the analysis of WBM in commercial meatball samples.

Matched MeSH terms: 1-Propanol
Fulltext Determination of volatile compounds in fresh and fermented Nipa sap (Nypa fruticans) using static headspace gas chromatography-mass spectrometry (GC-MS)

Nur Aimi, R., Abu Bakar, F., Dzulkifly, M.H.

International Food Research Journal, 2013;20(1):369-376.
MyJurnal

Nipa sap or air nira is a sweet natural beverage obtained from a type of palm tree, Nypa fruticans.
It is readily and spontaneously fermented resulting in the development of alcoholic fermentation products. Objective of this study is to determine the volatile compounds (VOCs) responsible for the aroma in fresh and fermented nipa sap. The sap was left for natural fermentation at 30ºC for 63 days. VOCs of the sap were analysed using static headspace gas chromatography-mass spectrometry (GC-MS). Fresh nipa sap contained ethanol (83.43%), diacetyl (0.59%), and esters
(15.97%). Fermented nipa sap contained alcohols (91.16 – 98.29%), esters (1.18 – 8.14%), acetoin (0.02 – 0.7%), diacetyl (0.04 – 0.06%), and acetic acid (0.13 – 0.68%). Concentration of ethanol in fresh nipa sap increased from 0.11% (v/v) to 6.63% (v/v) during the fermentation, and slightly decreased to 5.73% (v/v) at day 63. No higher alcohols were detected in the fresh nipa sap. Concentration of 1-propanol and 2-methylpropanol were constant throughout the fermentation with average of 0.004 to 0.006% (v/v) and 0.0001 to 0.0009% (v/v), respectively. 3-methylbutanol increased during the fermentation process. The highest concentration (0.001% v/v) was recorded at day 35. This study has shown differences in VOCs types between fresh and fermented nipa sap.

Matched MeSH terms: 1-Propanol
Fulltext Lid opening and conformational stability of T1 Lipase is mediated by increasing chain length polar solvents

Maiangwa J, Mohamad Ali MS, Salleh AB, Rahman RNZRA, Normi YM, Mohd Shariff F, et al.

PeerJ, 2017;5:e3341.
PMID: 28533982 DOI: 10.7717/peerj.3341

The dynamics and conformational landscape of proteins in organic solvents are events of potential interest in nonaqueous process catalysis. Conformational changes, folding transitions, and stability often correspond to structural rearrangements that alter contacts between solvent molecules and amino acid residues. However, in nonaqueous enzymology, organic solvents limit stability and further application of proteins. In the present study, molecular dynamics (MD) of a thermostable Geobacillus zalihae T1 lipase was performed in different chain length polar organic solvents (methanol, ethanol, propanol, butanol, and pentanol) and water mixture systems to a concentration of 50%. On the basis of the MD results, the structural deviations of the backbone atoms elucidated the dynamic effects of water/organic solvent mixtures on the equilibrium state of the protein simulations in decreasing solvent polarity. The results show that the solvent mixture gives rise to deviations in enzyme structure from the native one simulated in water. The drop in the flexibility in H2O, MtOH, EtOH and PrOH simulation mixtures shows that greater motions of residues were influenced in BtOH and PtOH simulation mixtures. Comparing the root mean square fluctuations value with the accessible solvent area (SASA) for every residue showed an almost correspondingly high SASA value of residues to high flexibility and low SASA value to low flexibility. The study further revealed that the organic solvents influenced the formation of more hydrogen bonds in MtOH, EtOH and PrOH and thus, it is assumed that increased intraprotein hydrogen bonding is ultimately correlated to the stability of the protein. However, the solvent accessibility analysis showed that in all solvent systems, hydrophobic residues were exposed and polar residues tended to be buried away from the solvent. Distance variation of the tetrahedral intermediate packing of the active pocket was not conserved in organic solvent systems, which could lead to weaknesses in the catalytic H-bond network and most likely a drop in catalytic activity. The conformational variation of the lid domain caused by the solvent molecules influenced its gradual opening. Formation of additional hydrogen bonds and hydrophobic interactions indicates that the contribution of the cooperative network of interactions could retain the stability of the protein in some solvent systems. Time-correlated atomic motions were used to characterize the correlations between the motions of the atoms from atomic coordinates. The resulting cross-correlation map revealed that the organic solvent mixtures performed functional, concerted, correlated motions in regions of residues of the lid domain to other residues. These observations suggest that varying lengths of polar organic solvents play a significant role in introducing dynamic conformational diversity in proteins in a decreasing order of polarity.

Matched MeSH terms: 1-Propanol
Fulltext Comparison of hydro-distillation, hydro-distillation with enzyme-assisted and supercritical fluid for the extraction of essential oil from pineapple peels

Mohamad N, Ramli N, Abd-Aziz S, Ibrahim MF

3 Biotech, 2019 Jun;9(6):234.
PMID: 31139549 DOI: 10.1007/s13205-019-1767-8

Pineapple peel is a potential feedstock for the extraction of essential oil due to the presence of aromatic compounds. To extract the essential oil from pineapple peels, three different methods were applied, i.e., (1) hydro-distillation (HD); (2) hydro-distillation with enzyme-assisted (HDEA); and (3) supercritical fluid extraction (SFE). SFE had successfully produced an essential oil with the yield of 0.17% (w/w) together with 0.64% (w/w) of concrete, whereby the HD and HDEA had only produced hydrosols with the yield of 70.65% (w/w) and 80.65% (w/w), respectively. Parameters' optimization for HD (substrate to solvent ratio, temperature, and extraction duration) and HDEA (cellulase loading and incubation duration) significantly affected the hydrosol yield, but did not extract out the essential oil. This is because only SFE had successfully ruptured the oil gland after observed under the scanning electron microscope. The essential oil obtained from SFE composed of mainly propanoic acid ethyl ester (40.25%), lactic acid ethyl ester (19.35%), 2-heptanol (15.02%), propanol (8.18%), 3-hexanone (2.60%), and butanoic acid ethyl ester (1.58%). In overall, it can be concluded that the SFE had successfully extracted the essential oil as compared to the HD and HDEA methods.

Matched MeSH terms: 1-Propanol
Determination of diffusion coefficients and activation energy of selected organic liquids using reversed-flow gas chromatographic technique

Khalisanni Khalid, Rashid Atta Khan, Sharifuddin Mohd. Zain

Sains Malaysiana, 2012;41:1109-1116.

Evaporation of vaporize organic liquid has ecological consequences when the compounds are introduced into both freshwater and marine environments through industrial effluents, or introduced directly into the air from industrial unit processes such as bioreactors and cooling towers. In such cases, a rapid and simple method are needed to measure physicochemical properties of the organic liquids. The Reversed-Flow Gas Chromatography (RF-GC) sampling technique is an easy, fast and accurate procedure. It was used to measure the diffusion coefficients of vapors from liquid into a carrier gas and at the same time to determine the rate coefficients for the evaporation of the respective liquid. The mathematical expression describing the elution curves of the samples peaks was derived and used to calculate the respective parameters for the selected liquid pollutants selected such as methanol, ethanol, 1-propanol, 1-butanol, n-pentane, n-hexane, n-heptane and n-hexadecane, evaporating into the carrier gas of nitrogen. The values of diffusion coefficients found were compared
with those calculated theoretically or reported in the literature. The values of evaporation rate were used to determine the activation energy of respective samples using Arrhenius equation. An interesting finding of this work is by using an alternative mathematical analysis based on equilibrium at the liquid-gas interphase, the comparison leads to profound
agreement between theoretical values of diffusion coefficients and experimental evidence.

Matched MeSH terms: 1-Propanol