Oumaima Maroua Bouguerra, Roswanira Abdul Wahab, Fahrul Huyop, Naji Arafat Mahat, Evi Susanti
Despite extensive characterization of its biochemical properties, Acinetobacter haemolyticus recombinant alkaline-stable lipase KV1 (Lip-KV1) remains unexplored in crosslinked enzyme aggregates (CLEAs) as a means to enhance its catalytic performance. This research focused on optimizing the preparation of Lip-KV1CLEAs through the application of response surface methodology and Box–Behnken design (BBD) to optimize the parameters, glutaraldehyde concentration (mM), Tween 80 (v/v%), pH, and temperature, for the response of recovery activity (%). Results revealed that the CLEAs-Lip-KV1 exhibited the highest recovery activity of 123.3% under the following optimum condition: isopropanol as the precipitant (80% v/v), Tween 80 at 0.01% (v/v), 60 mM glutaraldehyde, agitation at 200 rpm, pH 4.0 with a 2-h crosslinking duration at 30 °C. The resultant CLEAs-Lip-KV1 was stable in 10% v/v of methanol. Notably, the optimum temperature and pH stability of the CLEAs-Lip-KV1 was elevated compared to its free form, from the corresponding 40 °C and pH 8, to 50 °C and pH 10, with its thermal stability reaching up to 80 °C. The FTIR spectrum and FESEM micrographs confirmed these changes chemically and morphologically, which verified the CLEAs-Lip-KV1 preparation. © King Fahd University of Petroleum & Minerals 2025.
Department of Bioscience, Faculty of Science, Universiti Teknologi Malaysia, UTM, Johor, Johor Bahru, 81310, Malaysia; Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, UTM, Johor, Johor Bahru, 81310, Malaysia; Advanced Membrane Technology Research Centre (AMTEC), Universiti Teknologi Malaysia, UTM, Johor Bahru, 81310, Malaysia; Investigative and Forensic Science Research Group, Faculty of Science, Universiti Teknologi Malaysia, Johor, Johor Bahru, 81310, Malaysia; Centre for Sustainable Nanomaterials, Ibnu Sina Institute for Scientific and Industrial Research, Universiti Teknologi Malaysia, Johor, Johor Bahru, 81310, Malaysia; Faculty of Syariah and Law, Centre of Research for Fiqh Forensics and Judiciary, Universiti Sains Islam, Nilai, Malaysia; Biotechnology Program, Department of Applied Science, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Malang, Indonesia
