Production of clovamide and its analogues in Saccharomyces cerevisiae and Lactococcus lactis.

Clovamide and its analogues are N-hydroxycinnamoyl-L-amino acids (HAA) that exhibit antioxidant activities. For environmental and economic reasons, biological synthesis of these plant-derived metabolites has garnered interest. In this study, we exploited HDT1, a BAHD acyltransferase recently isolated from red clover, for the production of clovamide and derivatives in S. cerevisiae and L. lactis. HDT1 catalyses the transfer of hydroxycinnamoyl-coenzyme A (CoA) onto aromatic amino acids. Therefore, by heterologously co-expressing HDT1 with 4-coumarate:CoA ligase (4CL), we succeeded in the biological production of clovamide and more than 20 other HAA, including halogenated ones, upon feeding the engineered micro-organisms with various combinations of cinnamates and amino acids. To the best of our knowledge, this is the first report on the biological synthesis of HAA and, more generally, on the synthesis of plant-derived antioxidant phenolic compounds in L. lactis. The production of these health beneficial metabolites in Generally Recognized As Safe (GRAS) micro-organisms such as S. cerevisiae and L. lactis provides new options for their delivery as therapeutics. SIGNIFICANCE AND IMPACT OF THE STUDY: N-hydroxycinnamoyl-L-amino acids such as clovamide are bioactive plant-derived phenolic compounds with health beneficial effects. Relying on chemical synthesis or direct extraction from plant sources for the supply of these valuable molecules poses challenges to environmental sustainability. As an alternative route, this work demonstrates the potential for biological synthesis of N-hydroxycinnamoyl-L-amino acids using engineered microbial hosts such as Saccharomyces cerevisiae and Lactococcus lactis. Besides being more eco-friendly, this approach should also provide more structurally diverse compounds and offer new methods for their delivery to the human body.

Menarche and menstrual function in patients with myelomeningocele.

The aim of this study was to assess menarchal age, to clarify the role of heredity in the timing of menarche, and to examine the management of menses in girls and young women with myelomeningocele. All female patients aged three years or older attending a tertiary-care hospital clinic were eligible. A questionnaire was administered to each menstruating patient and, when possible, a menstruating sibling control. Most biological descriptors of menses were comparable for patients and sibling controls, but management of menses, routine gynecological care and attitudes about family planning differed. Girls with myelomeningocele menstruated significantly earlier than the US mean (10.25 years); as did the girls' mothers (11.91 years), but not their sisters. This finding may have biological significance, and a larger sample size is needed to confirm or refute it.