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Supporting InformationThe Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/jasms.9b00074.Step-by-step protocol and important notes for optimizing the HTX TM-spraying parameters using the RSM, containing (Table S1) coded and actual values of factors used in the experimental design, (Figure S1) organization of tissue biopsy sections on slides together with HTX TM-sprayer conditions used for each section, (Figure S2) example METASPACE webpage that shows the uploading process, (Figure S3) example METASPACE webpage that displays putative molecular annotations and corresponding ion images, and (Figure S4) examples of endogenous and off-tissue annotations by METASPACE (PDF)The authors declare no competing financial interest.

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Olive oil cake is a by-product from the olive oil processing industry and can be used for the lipase and protease production by Candida utilis in solid state fermentation. Different carbon and nitrogen sources were evaluated, and the results showed that the supplementation of the substrate with maltose and starch as carbon sources and yeast extract as a nitrogen source significantly increased the lipase production. The best results were obtained with maltose, whereas rather low lipase and protease activities were found with glucose and oleic acid. Response surface methodology and a five-level-three-factor central composite rotatable design were used to evaluate the effects of the initial moisture content, inoculum size and fermentation time on both lipase and protease activity levels. A lipase activity value of ≈25 U g(-1) and a protease activity value of 110 U g(-1) were obtained under the optimized fermentation conditions. An alkaline treatment of the substrate appeared to be efficient, leading to increases of 39% and 133% in the lipase and protease production, respectively. The results showed that the olive cake could be a good source for enzyme production by solid state fermentation.

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