Lycopene

Biweight midcorrelation, an outlier-robust correlation measure, was used to assess marginal linear relationships between epigenetic aging measures and dietary, cardiometabolic, and socioeconomic factors. To adjust for possible socioeconomic and lifestyle confounders, we fit ethnically-stratified multivariable linear models adjusting for education, exercise, BMI, and current drinker and smoker status. We used Stouffer's method to infer the meta-analytic significance of each variable over the different ethnic strata using the square-root of the sample size as the Z-score weighting factor. Specifically for the WHI, the age acceleration measures were adjusted for differences in originating dataset and within the InCHIANTI the measures were adjusted for sex. Models including regression on biomarkers, and number of metabolic syndrome symptoms were not stratified by ethnicity due to lack of coverage for biomarker profiling. Models were designed based on common prior knowledge and in cases where there was co-linearity between confounding variables, choice for adjustment was selected based on variable commonality in order to improve comparability with other studies, e.g. BMI was chosen over WHR because BMI is more commonly measured and reported. Variables with skewness >1 were log transformed (possibly adding +1 to avoid forming the logarithm of zero). Mean carotenoids was computed as the mean across standardized measures of lycopene, log2(alpha-carotene), log2(beta-carotene), log2(lutein + zeaxanthin), and log2(beta-cryptoxanthin). Repeat measurements on the same individuals were omitted from the analysis.

Many fruits and vegetable waste acts as DF. This waste, at the same time, contains such compounds, which show antioxidant properties. In addition to protecting cells from damage caused by free radicals, chemicals like polyphenols can also treat diseases and their symptoms by inhibiting inflammatory responses and halting the progression of infection (Shay et al., 2015 (link)). As a result, incorporating such substances into meat products may improve their functionality and, thus, their healthfulness. The remnants of fresh dates are the source of polyphenols; when they were included in the formulation of bologna sausages (15% of the total), the finished product had a polyphenol level of 1.02% (Sánchez-Zapata et al., 2011 (link)). This finding suggests that adding extracts rich in polyphenolic chemicals to meat products can serve as an antioxidant and provide health benefits to the end user. Lycopene, a carotenoid present in 80-90% of ripe tomatoes, has been linked to various health benefits, including a reduced risk of prostate cancer and CV disease (Friedman, 2013 (link)). After 21 days in storage, 0.58 mg of lycopene per 100 g of product was discovered in concentrations of up to 1.2% of the tomato peel in sausage. Lycopene was detected in beef burgers cooked at 180 °C for 2 min (Luisa García, Calvo & Dolores Selgas, 2009 (link)). Furthermore, the leftovers from tomatoes can be a source of amino acids and trace elements. By incorporating just 7% of the residue, the protein level was raised by 1%, while also boosting the ash content from 2.18% to 2.45% in frankfurter beef sausages. The percentage of total lipids dropped from 20.07 to 19.4 as a result (Savadkoohi et al., 2014 (link)).
Prebiotics are a potential health benefit of fruit and vegetable waste. Prebiotics are elements that can survive stomach acid, mammalian enzymatic hydrolysis, absorption in GI tract; they are fermentable by intestinal flora and hence foster the expansion of beneficial bacteria like probiotics (Gibson et al., 2004 (link)). Prebiotics come in many forms, but some common ones include cellulose and fiber. Fiber from nopal flour (2%) and pineapple peel flour (3%) added to cooked sausages helped inoculated thermos-tolerant (probiotic) lactic acid bacteria thrive over 20 days in storage (Díaz-Vela, Totosaus & Pérez-Chabela, 2015 (link)). It is important to note that the amount of bacteria in a formulation such as this one, which contains both a probiotic and a prebiotic, needs to be closely controlled because the bacteria have the potential to degrade the overall quality of the product. It has also been observed in the above-discussed case that the inclusion of agro-industrial waste raises the mineral content of the meat products, which could lead to a rise in mineral consumption and help meet dietary guidelines.

L-lactate dehydrogenase (LlLDH) was from Lactococcus lactis NZ9000. Acetolactate synthase (cytoILV2) was native S. cerevisiae gene but was truncated (2–90 aa) [8 (link)] to relocate it to the cytoplasm. Both LlLDH and cytoILV2 were inserted into plasmid pSP-GM2 between the restriction sites BamHI and NheI to construct plasmids used for production of lactate and 2,3-BD, respectively. Malonyl-CoA reductase (CaMCR), α-farnesene synthase (MdFS) and tyrosine ammonia lyase (FjTAL), from Chloroflexus aurantiacus [46 (link)], Malus domestica [61 (link)] and Flavobacterium johnsoniae [62 (link)], respectively, were codon optimized and placed under the control of TEF1 promoter, with their expression cassettes subsequently integrated into chromosomal site XII-2 to construct strains producing 3-HP, farnesene and p-coumaric acid. Similarly, geranylgeranyl pyrophosphate synthase (PaCrtE), phytoene synthase (PaCrtB) and phytoene desaturase (PaCrtI), all from Pantoea ananatis, were codon optimized, with their expression placed under the control of the promoters CDC19, CCW12 and TDH3, respectively. As before, the expression cassettes were then integrated into chromosomal site XII-2 to construct lycopene-producing strains. FFAs-producing strains were constructed by deleting FAA1, FAA4, POX1 and PAH1.
Expression formats were chosen according to previous study. As reported, 2,3-BD [16 (link), 63 (link)–65 (link)] and lactate [20 (link), 66 (link)–68 (link)] were widely produced by expressing genes in plasmids; meanwhile, p-coumaric acid [21 (link), 29 (link), 69 (link)–71 (link)], lycopene [44 (link), 45 (link), 72 (link), 73 (link)], farnesene [3 (link), 43 (link), 74 (link)] and 3-HP [75 (link)] were widely produced by integrating genes into genome. Using the same expression format as reported will be of referential significance for demonstrating the biosynthetic capacity of chassis cell.
All the plasmids and expression cassettes were transformed into yeast by LiAC/ssDNA method [76 (link)]. Expression cassettes were integrated by CRISPR/Cas9 system. All strains used in this study are listed in Table 1. All primers, gRNA and deleting donors used in this study were summarized in Additional file 1: Tables S1 and S2.

3-HP and lactate were measured at 65 °C by an HPLC system equipped with a refractive-index detector and a Bio-Rad HPX-87H column using 0.5 mM H₂SO₄ as mobile phase at a flow rate of 0.6 mL min⁻¹. Esterification and analysis of FFAs were performed as previously described [78 (link)]. 2,3-BD was detected by the same methods as 3-HP, except for the use of 5 mM H₂SO₄ as the mobile phase and column temperature of 28 °C. Lycopene and farnesene were detected by HPLC equipped with an Innoval C18 column. The extracted and analyzed method was described previously [45 (link)]. p-Coumaric acid was detected by HPLC equipped with Discovery HS F5 150 mm × 2.1 mm column, as described in a previous study [62 (link)].
All data were presented as the mean of three biological replicates with error bars representing the standard deviations. P values were generated by performing t tests for determining statistical significance.