Saisavoey Tanatorn, Sangtanoo Papassara, Srimongkol Piroonporn, Reamtong Onrapak, Karnchanatat Aphichart
Research Unit in Bioconversion/Bioseparation for Value-Added Chemical Production, Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, 254 Phayathai Road, Pathumwan, Bangkok, 10330 Thailand.
Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok, 10400 Thailand.
J Food Sci Technol. 2021 Feb;58(2):752-763. doi: 10.1007/s13197-020-04592-2. Epub 2020 Jun 23.
In order to examine bee pollen hydrolysates to assess their anticancer and antioxidant properties, hydrolysis of bee pollen was first performed using three different commercially available enzymes: Alcalase, Neutrase, and Flavourzyme. The study used DPPH and ABTS assay to evaluate the antioxidant properties of the hydrolysates obtained from bee pollen. All of the tested hydrolysates demonstrated antioxidant activity, while hydrolysate based on Alcalase offered a high value for IC and was therefore chosen for further separation into five sub-fractions via ultrafiltration. The greatest antioxidant activity was presented by the MW < 0.65 kDa fraction, which achieved an IC value of 0.39 ± 0.01 µg/mL in the DPPH assay and 1.52 ± 0.01 µg/mL for ABTS. Purification of the MW < 0.65 kDa fraction was completed using RP-HPLC, whereupon the three fractions from the original six which had the highest antioxidant activity underwent further examination through ESI-Q-TOF-MS/MS. These particular peptides had between 7 and 11 amino acid residues. In the case of the MW < 0.65 kDa fraction, testing was also carried out to determine the viability of lung cancer cell lines, represented by ChaGo-K1 cells. Analysis of the antiproliferative properties allowed in vitro assessment of the ChaGo-K1 cells' viability following treatment using the MW < 0.65 kDa fraction. Flow-cytometry generated date which revealed that it was possible for the MW < 0.65 kDa fraction to induce apoptosis in the ChaGo-K1 cells in comparison to the results with cells which had not been treated.
为了检测蜂花粉水解产物以评估其抗癌和抗氧化特性,首先使用三种不同的市售酶:碱性蛋白酶、中性蛋白酶和风味酶对蜂花粉进行水解。该研究使用DPPH和ABTS测定法评估从蜂花粉获得的水解产物的抗氧化特性。所有测试的水解产物都表现出抗氧化活性,而基于碱性蛋白酶的水解产物的IC值较高,因此被选择通过超滤进一步分离成五个亚组分。MW < 0.65 kDa组分表现出最大的抗氧化活性,在DPPH测定中其IC值达到0.39 ± 0.01 µg/mL,在ABTS测定中为1.52 ± 0.01 µg/mL。使用反相高效液相色谱法(RP-HPLC)完成了MW < 0.65 kDa组分的纯化,随后对原始六个具有最高抗氧化活性的组分中的三个进行了电喷雾四极杆飞行时间串联质谱(ESI-Q-TOF-MS/MS)进一步检测。这些特定的肽含有7至11个氨基酸残基。对于MW < 0.65 kDa组分,还进行了测试以确定以ChaGo-K1细胞为代表的肺癌细胞系的活力。对其抗增殖特性的分析允许在使用MW < 0.65 kDa组分处理后对ChaGo-K1细胞的活力进行体外评估。流式细胞术产生的数据表明,与未处理的细胞相比,MW < 0.65 kDa组分有可能诱导ChaGo-K1细胞凋亡。