{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:45:43Z","timestamp":1768509943182,"version":"3.49.0"},"posted":{"date-parts":[[2024,1,1]]},"group-title":"Immunology","reference-count":35,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,4,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                  is a fungal pathogen that causes cryptococcosis mostly in immune compromised patients, such as those with HIV\/AIDS. One survival mechanism of\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  during infection is melanin production, which catalyzed by laccase, and protects fungal cells against immune attack. Hence comparative assessment of laccase activity is useful for characterizing cryptococcal strains. We serendipitously observed that culturing\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  with food coloring resulted in the degradation of some dyes with phenolic structures. Consequently, we investigated the color changes for the food dyes metabolized by\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  laccase and explored using this effect for the development of a colorimetric assay to measure laccase activity. We developed several versions of a food dye based colorimetric laccase assay that can be used to compare the relative laccase activities between different\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  strains. We found that phenolic color degradation was glucose dependent, which may reflect changes in the reduction properties of the media. Our food color based colorimetric assay has several advantages over the commonly used 2,2\u2032-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay for laccase activity, including lower cost, irreversibility, and does not require constant monitoring. This method has potential applications to bioremediation of water pollutants in addition to its use in determining laccase virulence factor expression.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                    is present in the environment and while infection is common, disease occurs mostly in immunocompromised individuals.\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    infection in the lungs results in symptoms like pneumonia, and consequently cryptococcal meningitis occurs if the fungal infection spreads to the brain. The laccase enzyme catalyzes the melanization reaction that serves as a virulence factor of\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    . Developing a simple and less costly assay to determine laccase activity in\n                    <jats:italic>C. neoformans<\/jats:italic>\n                    strains can be useful for a variety of procedures ranging from studying the relative virulence of cryptococci to environmental pollution studies.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.01.01.573823","type":"posted-content","created":{"date-parts":[[2024,1,1]],"date-time":"2024-01-01T22:00:11Z","timestamp":1704146411000},"source":"Crossref","is-referenced-by-count":1,"title":["A food color based colorimetric assay for\n                  <i>Cryptococcus neoformans<\/i>\n                  laccase activity"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4110-9873","authenticated-orcid":false,"given":"Lia Sanchez","family":"Ramirez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4068-5828","authenticated-orcid":false,"given":"Quigly","family":"Dragotakes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9402-9167","authenticated-orcid":false,"given":"Arturo","family":"Casadevall","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024041002550981000_2024.01.01.573823v2.1","unstructured":"C. neoformans Infection Statistics | Fungal Diseases | CDC. https:\/\/www.cdc.gov\/fungal\/diseases\/cryptococcosis-neoformans\/statistics.html (2022)."},{"key":"2024041002550981000_2024.01.01.573823v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsyr.2004.04.004"},{"key":"2024041002550981000_2024.01.01.573823v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(00)00103-X"},{"key":"2024041002550981000_2024.01.01.573823v2.4","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2008.06245.x"},{"key":"2024041002550981000_2024.01.01.573823v2.5","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1128\/jcm.7.2.146-152.1978","article-title":"Pigment production by Cryptococcus neoformans and other Cryptococcus species from aminophenols and diaminobenzenes","volume":"7","year":"1978","journal-title":"J. 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