In this work we report the synthesis of organic-inorganic solid with spherical morphology where enzyme, as active compounds, is encapsulated. The organic phase of nanospheres is composed of L-α-phosphatidylcholine, as liposome, and Lipase from Rhizomucor miehei, as enzyme. The organic phase is covered with porous inorganic silica shell that could stabilize the internal liposomal phase and, consequently, isolate and protect the bioactive molecules. The liposome and silica amount used during the immobilization procedure have been optimized in order to obtain active and stable heterogeneous biocatalyst. Hybrid-nanospheres containing the enzyme were used to catalyze the transesterification reaction of triolein with methanol to methyl esters, typical biodiesel mixture compounds. The encapsulated enzyme retains its activity after 5 reaction cycles. The total productivity of the best catalyst obtained is higher than that of the free enzyme.
Answer to CommentsReviewer #1: The authors in the article described a new method of preparation of encapsulated active biomolecules. The lipase enzymes in silica nanoparticles are catalytical more active for the transesterification reaction than free enzymes, even after 5 reaction cycles. I recommend the article for publication in Catalysis Today.A: We want to thank the reviewer for the revision.Reviewer #2: The paper deals with an interesting topic and merits to be published in Catalysis Today. However, the following comments should be taken into account prior to final acceptance of the manuscript:A: We have appreciated very much all the suggestions made by the reviewer. They have entailed important improvements of the manuscript.
Main comments:Q1: What is the productivity compared with classical homogeneous catalysts and some heterogeneous catalysts reported in literature? Please provide a comparison.A1: In order to make this comparison, we have introduced another paragraph, "Comparison with some homogeneous and heterogeneous catalysts", where we have compared the best performance of some heterogeneous catalysts and the classical homogeneous catalysts (basic, KOH, and acid, H 2 SO 4 ). Unfortunately, we don't have the productivity values for all these catalysts and, in any case, the tested enzyme amount and the number of reaction cycles should be the same, in order to make a good comparison. Therefore we have compared the overall catalytic performance. To do this comparison, we have inserted some references (from 15 to 19). Q2: What is the reason for the rapid drop in activity after the fifth cycle? Please explain it in the text.A2: Most probably, the main reason of the rapid drop of the enzyme activity after the fifth cycle should be the denaturation of the same enzyme, together with the inevitable leaching. We have introduced this consideration in the text, section 3.3 Transesterification reaction results, lines 54-55.
*Detailed Response to ReviewersQ3: It is said: "The possibility to use cheaper feedstock requires alternative catalysts instead of the homogeneous basi...