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The authors describe an aptamer based assay for the food mycotoxin ochratoxin A (OTA). It is based on the use of exonuclease III (Exo III) which assists in signal amplification, and of single-walled carbon nanohorns (SWCNHs) which act as quenchers of fluorescence. The detection scheme employs a hairpin probe (HP) and a signal probe (SP) labeled with carboxyfluorescein (FAM) at its 5'-end. The fluorescence of intact SPs (best measured at excitation/emission wavelengths of 495/518 nm) is quenched by SWCNHs. The HP contains the OTA-specific aptamer sequence and is partially complementary to the SP. After addition of OTA, the aptamer binds OTA and thus exposes a single-stranded sequence that can hybridize with the SP. Exo III digests the SP to liberate the free fluorophore labels. The damaged SPs no longer are adsorbed by the SWCNHs so that fluorescence is no longer quenched. The method has a detection range that is linear from 10 nM to 1000 nM (with a correlation coefficient of 0.997). The limit of detection (LOD), calculated on the basis of a signal to noise ratio of 3, is 4.2 nM. The procedure was validated by the quantitation of OTA in spiked real samples and were found to be free of interference by the sample matrix. Recoveries ranged from 93.8 to 113.0% in beer and from 92.0 to115.9% in red wine. Graphical abstract After adding ochratoxin A (OTA), the aptamer region in hairpin probe (HP) combined with OTA and thus exposed a single-stranded sequence to hybridize with signal probe (SP). Exonuclease III (Exo III) digested SP to liberate the free fluorophore (FAM).

An protease from for improving meat tenderness was purified, characterized from wild fruit by ammonium sulfate precipitation, Sephdex G-25 gel filtration chromatography, and DEAE Sepharose Fast Flow ion exchange chromatography, and its activity was investigated. The purified protease was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis to obtain a single band of protease. The protease was purified successfully, and found to have a molecular weight of 23.8 kDa (mass spectrometry). The specific activity of the purified protease reached 53,428 U/mg with a 25.5-fold purification factor and 9% activity recovery. Based on N-terminal sequencing results, the protease was derived from the class of actinidia proteases that have an N-terminal sequence of VLPDY VDWRS AGAVV. The protease was effective for tenderizing beef and decomposing actomyosin, suggesting the potential application for improving meat tenderness.

Introduction According to the different melting point, palm oil can be divided into palm stearin 44-56 , intermediate component 26-43 and palmitin 24. After being fractionated by acetone and other organic solvents, palm stearin can be obtained with high-content tripalmitin PPP of triglyceride. Tripalmitin and oleic acid methyl oleate, methyl oleate can be used to synthesize 1,3-dioleoyl-2-palmitoylglycerol OPO structured lipid by enzymatic acidolysis reaction 1, 2. OPO could be used as human milk fat substitute HMFS. And that human milk was a natural emulsion system 3. Human milk fat HMF was the richest energy source for infants, supplying about 50-60 of dietary calories. Approximately 98 of the HMF was in the form of triacylglycerol TAG with a unique fatty acid tribution. Dietary TAGs are hydrolyzed into 2-monoacylglycerol 2-MAG and free fatty acids by pancreatic lipase in the intestine.