Gallic acid-based alkyl esters synthesis in a water-free system by celite-bound lipase of Bacillus licheniformis SCD11501.
Gallic acid (3, 4, 5- trihydroxybenzoic acid) is an important antioxidant, anti-inflammatory, and radical scavenging agent. In the present study, a purified thermo-tolerant extra-cellular lipase of Bacillus licheniformis SCD11501 was successfully immobilized by adsorption on Celite 545 gel matrix followed by treatment with a cross-linking agent, glutaraldehyde. The celite-bound lipase treated with glutaraldehyde showed 94.8% binding/retention of enzyme activity (36 U/g; specific activity 16.8 U/g matrix; relative increase in enzyme activity 64.7%) while untreated matrix resulted in 88.1% binding/retention (28.0 U/g matrix; specific activity 8.5 U/g matrix) of lipase. The celite-bound lipase was successfully used to synthesis methyl gallate (58.2%), ethyl gallate (66.9%), n-propyl gallate (72.1%), and n-butyl gallate (63.8%) at 55(o) C in 10 h under shaking (150 g) in a water-free system by sequentially optimizing various reaction parameters. The low conversion of more polar alcohols such as methanol and ethanol into their respective gallate esters might be due to the ability of these alcohols to severely remove water from the protein hydration shell, leading to enzyme inactivation. Molecular sieves added to the reaction mixture resulted in enhanced yield of the alkyl ester(s). The characterization of synthesised esters was done through fourier transform infrared (FTIR) spectroscopy and (1) H NMR spectrum analysis.
Dept. of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, 171 005, India.
Shivika
S
Sharma
Dept. of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, 171 005, India.
Shamsher S
SS
Kanwar
Dept. of Chemistry, Himachal Pradesh University, Summer Hill, Shimla, 171 005, India.
Priyanka
P
Dogra
Dept. of Chemistry, Himachal Pradesh University, Summer Hill, Shimla, 171 005, India.
Ghanshyam S
GS
Chauhan
D001407:Bacillus / Q000201:enzymology*; D007692:Diatomaceous Earth / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000378:metabolism; D004951:Esterification; D004952:Esters; D000431:Ethanol / Q000378:metabolism; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000378:metabolism*; D008049:Lipase / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000378:metabolism; D011435:Propyl Gallate / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014867:Water
D004800:Enzymes, Immobilized; D004952:Esters; D014867:Water; C048734:ethyl gallate; D000431:Ethanol; D007692:Diatomaceous Earth; C052082:methyl gallate; D005707:Gallic Acid; C015048:n-butyl gallate; D011435:Propyl Gallate; D008049:Lipase; D000432:Methanol
butanol; esterification; ethanol; gallic acid; immobilization; lipase; methanol; propanol