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D'Amore, L.

Publications and source records attributed to D'Amore, L..

2 recordsLinked to original sources

B-vitamin requirements and phycosphere interactions for the HAB-forming dinoflagellate, Pyrodinium bahamense var. bahamense

Pyrodinium bahamense is a saxitoxin-producing dinoflagellate found in tropical and subtropical estuarine waters globally, with the var. bahamense variety forming recurrent harmful algal blooms (HABs) in Florida estuaries. The intensity and duration of bloom peaks in Florida estuaries vary interannually, but life cycle transitions, estuarine water residence time, water temperature, salinity, and nutrient concentrations appear to contribute to bloom formation. However, the impact of B-vitamin availability on P. bahamense bloom formation has not been investigated, despite the knowledge that many HAB dinoflagellates require B-vitamins for growth and bloom development. In this study, we determined that of the three B-vitamins most commonly required by marine phytoplankton--biotin (B7), thiamine (B1), and cobalamin (B12)--P. bahamense strictly requires an exogenous B12 source to sustain growth. As bacterial interactions can fulfill phytoplankton B12 requirements, we tested whether bacterial communities recruited from two Florida estuaries could rescue P. bahamense from B12 limitation. We demonstrate that bacteria from the Indian River Lagoon (IRL) estuary collected during a P. bahamense bloom restored P. bahamense growth in media missing B12, while bacteria present in Old Tampa Bay (OTB) water collected when P. bahamense was not present did not restore P. bahamense growth in media missing B12. Putative bacterial B12 producers exclusively recruited from IRL included Enterobacteraceae genera that likely have anthropogenic sources. Our findings suggest B12 is essential for fueling P. bahamense blooms, and that specific bacterial communities may contribute to bloom development through B12 production.

microbiology↗

Pervasive cooperative mutational effects on multiple catalytic enzyme traits emerge via long-range conformational dynamics

Multidimensional fitness landscapes provide insights into the molecular basis of laboratory and natural evolution. Yet such efforts are rare and focus only on limited protein families and a single enzyme trait, with little concern about the relationship between protein epistasis and conformational dynamics. Here, we report the first multiparametric fitness landscape for a cytochrome P450 monooxygenase that was engineered for the regio- and stereoselective hydroxylation of a steroid. We developed a computational program to automatically quantify non-additive effects among all possible mutational pathways, finding pervasive cooperative sign and magnitude epistasis on multiple catalytic traits. By using quantum mechanics and molecular dynamics simulations, we show that these effects are modulated by long-range interactions in loops, helices and beta-strands that gate the substrate access channel allowing for optimal catalysis. Our work highlights the importance of conformational dynamics on epistasis in an enzyme involved in secondary metabolism and offers lessons for engineering P450s.

evolutionary biology↗