The production of malolactic starter cultures requires the obtention of suitably large biomass at low-cost. In this work it was possible to obtain a good amount of biomass, at laboratory scale, of two enological strains of Lb. plantarum, by formulating a culture medium based on whey permeate (WP), a by-product of the cheese industry usually disposed as waste, when this was supplemented with yeast extract (Y), salts (S) andTween 80 (T) (WPYST). Bacteria grown in WPYST medium exhibited good tolerance to stress conditions of synthetic wine (pH 3.5, ethanol 13% vol/vol). However, when WPYST was added with 8% vol/vol ethanol, cultures inoculated in synthetic wine, showed a lower viability and capacity to consume L-malic acid than when they were cultured in WPYST without ethanol. Subsequently, strains grown in WPYST were inoculated in sterile wine samples (final stage of alcoholic fermentation) of the red varietals Merlot and Pinot noir, and incubated at laboratory scale. Cultures from WPYST, inoculated in Pinot noir wine, showed a better performance than bacteria grown in MRS broth, and exhibited a consumption of L-malic acid higher than 90%. However, cultures from WPYST or from MRS broth, inoculated in sterile Merlot wine, showed a lower survival. This study allowed the formulation of a low-cost culture medium, based on a by-product of the food industry, which showed to be adequate for the growth of two enological strains of Lb. plantarum, suggesting their potentiality for application in the elaboration of malolactic starter cultures. K E Y W O R D Senological Lactobacillus plantarum strains, malic acid consumption, nutritional supplementation, sterile wine, whey permeate

Circadian rhythms are endogenous oscillations present in nearly all organisms from prokaryotes to humans, allowing them to adapt to cyclical environments close to 24 hours. Circadian rhythms are regulated by a central clock, which is based on a transcription-translation feedback loop. One important protein in the central loop in metazoan clocks is PERIOD, which is regulated in part by Casein kinase 1ε/δ (CK1ε/δ) phosphorylation. In the nematodeCaenorhabditis elegans,periodandcasein kinase 1ε/δare conserved aslin-42andkin-20, respectively. Here we studied the involvement oflin-42andkin-20in circadian rhythms of the adult nematode using a bioluminescence-based circadian transcriptional reporter. We show that mutations oflin-42andkin-20generate a significantly longer endogenous period, suggesting a role for both genes in the nematode circadian clock, as in other organisms. These phenotypes can be partially rescued by overexpression of either gene under their native promoter. Both proteins are expressed in neurons and seam cells, a population of epidermal stem cells inC. elegansthat undergo multiple divisions during development. Depletion of LIN-42 and KIN-20 specifically in neuronal cells after development was sufficient to lengthen the period of oscillatingsur-5expression. Therefore, we conclude that LIN-42 and KIN-20 are critical regulators of the adult nematode circadian clock through neuronal cells.

The by-products of the food industry are an economic alternative as a source of nutrients to obtain biomass. At the same time, theiruse could solve the environmental problem related to their disposal, which is highly polluting due to their elevated biochemical oxygen demand. In this work, we seek to optimize the production of cellular biomass of two native Patagonian strains of Lactiplantibacillus plantarum (UNQLp 11 and UNQLp155), selected for its oenological and technological properties, using apple pomace (AP), a residue from the juice and cider industry. The supplementation of AP with yeast extract, salts, and Tween 80 (sAP), proved to maintain the growth of the Lpb. plantarum strains, similar to the commercial medium used to grow LAB (De Man, Rogosa, Sharpe, MRS). Cultures grown in sAP medium showed good tolerance to wine conditions (high ethanol content and low pH), demonstrated by its ability to consume L-malic acid. The subsequent inoculation of these cultures in sterile wines (Merlot and Pinot noir) was carried out at laboratory scale, evaluating cell viability and L-malic acid consumption for 21 days at 21 °C. Cultures grown in sAP media showed a similar performance to MRS media. Thus, sAP media proved to be a suitable substrate to grow oenological Lpb. plantarum strains where cultures (with high size inoculums) were able to drive malolactic fermentation, with an L-malic acid consumption higher than 90%.