Cottonseed Meal Protein Isolate as a New Source of Alternative Proteins: A Proteomics Perspective.

Cottonseed meal (CSM) is a good source of dietary proteins but is unsuitable for human consumption due to its gossypol content. To unlock its potential, we developed a protein extraction process with a gossypol removal treatment to generate CSM protein isolate (CSMPI) with ultra-low gossypol content. This process successfully reduced the free and total gossypol content to 4.8 ppm and 147.2 ppm, respectively, far below the US FDA limit. In addition, the functional characterisation of CSMPI revealed a better oil absorption capacity and water solubility than pea protein isolate. Proteome profiling showed that the treatment improved protein identification, while SDS-PAGE analysis indicated that the treatment did not induce protein degradation. Amino acid analysis revealed that post-treated CSMPI was rich in branched-chain amino acids (BCAAs). Mass spectrometry analysis of various protein fractions obtained from an in vitro digestibility assay helped to establish the digestibility profile of CSM proteins. Several potential allergens in CSMPI were also found using allergenic prediction software, but further evaluation based on their digestibility profiles and literature reviews suggests that the likelihood of CSMPI allergenicity remains low. Overall, our results help to navigate and direct the application of CSMPIs as alternative proteins toward nutritive human food application.

Micro-level evaluation of organic compounds transformation in anaerobic digestion under feast and famine conditions assisted by iron-based materials - Revealing the true mechanism of AD enhancement.

Conductive materials have been applied to assist syntrophic metabolism in anaerobic digestion. However, their role in the transformation of organic compounds, particularly recalcitrant compounds, has not been revealed. In this study, iron-based materials - magnetite nanoparticles and Fe2+- were employed to explore their effects on the transformation of different organic matters in anaerobic system. Prompted methane production rates and quantity in iron-based materials groups were found due to the improved solubilization of organic particles, enhanced degradation of recalcitrant compounds, and maintained microbial activity under substrate-limited conditions. Specifically, the proportion of the reducing functional groups (C-C/H or CC) and O/C ratio were always significantly lower in iron-based materials supplemented groups (Fe groups) compared to Control group, despite hydrolysis was greatly enhanced in Fe groups. The greater dehydrogenation oxidation was confirmed in the presence of iron-based materials. The remaining humic-like substances (HS), a typical type of recalcitrant compound, was about 2.5 times higher in Control group (221.2 ± 5.3 mg/L-C) compared to Fe groups after 30 days degradation. By tracking the aromaticity of HS and individual compounds at molecular level, this study reveals that iron-based materials were more effective in stimulating the degradation of aliphatic moieties than the aromatic moieties of recalcitrant compounds. When readily biodegradable substrates were limited, Fe groups continued methane generation by using recalcitrant compounds (e.g. thiethylperazine and fluvoxamino acid) as carbon source, and the microbial activity was maintained according to higher relative abundance of protonated nitrogen and continuous methanogenesis activity at starvation phase.

Biotransformation of phosphorus in enhanced biological phosphorus removal sludge biochar.

Biochar derived from enhanced biological phosphorus removal (EBPR) sludge could be a potential phosphorus (P) fertilizer. Soil microorganisms play a regulating role on the turnover of P in soil. When the EBPR sludge biochar is added to soil, it would inevitably interact with soil microorganisms. Thus, for the wise use of the EBPR sludge biochar, it is imperative to understand the interaction between the biochar and soil microorganisms. In this study, Pseudomonas putida (P. putida), a common soil microorganism, was applied to investigate the biotransformation of P in two EBPR sludge biochars. The results reveal that P released from biochar produced at 700 °C (E700) was more easily absorbed by P. putida than that released from biochar produced at 400 °C (E400). This is attributed to the higher polyphosphates (poly-P) content in E700 and poly-P has higher affinity to P. putida surface compared to orthophosphates. Furthermore, E400 has a negative effect on intracellular poly-P formation in P. putida, which is probably caused by the oxidative stress induced by the free radicals from E400. As intracellular poly-P plays a critical role on bacteria survival and their interaction with surrounding environment, high-temperature biochar (E700) in this case would be more suitable for soil remediation.