Ugyen Ugyen, Singanusong Raintong, Phinyo Mahattanee, Changtor Phanupong, Chaijamrus Sirilux, Thongsook Tipawan
Department of Agro-Industry, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, 65000 Phitsanulok, Thailand.
Department of Agricultural Science, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, 65000 Phitsanulok, Thailand.
Food Technol Biotechnol. 2023 Dec;61(4):523-535. doi: 10.17113/ftb.61.04.23.8255.
Heat-stabilised defatted rice bran (HSDRB) is a primary by-product of rice bran oil extraction industry and a nutritious source of protein. However, despite the unique nutritional profile of rice bran protein, the protein-rich by-product, HSDRB is underutilised as a low-value animal feed. Research on protein extraction from HSDRB by enzymatic hydrolysis has attracted the attention of numerous scientists. However, a cost-effective extraction method is required to mitigate the high costs associated with the use of enzymes. Therefore, we have presented an alternative economical and natural approach for protein extraction from HSDRB by solid-state fermentation (SSF) with heterofermentative microbes.
SSF of HSDRB with two types of traditional Asian fermentation starters, namely loog-pang and koji, were evaluated for enzyme production and their efficacy in extracting proteins from HSDRB. For this purpose, HSDRB fermentation was carried out for 0, 12, 24, 48, 72 and 96 h followed by 24-hour hydrolysis to evaluate the extracted rice bran protein. In addition, microbiome diversity in the fermentation starters was also determined by metagenomic sequencing of 16S rRNA and internal transcribed spacer to identify bacteria and fungi, respectively.
The microbial community in the fermentation starters showed the dominance of lactic acid bacteria (LAB) such as in loog-pang and , and in koji, while yeast species and dominated the fungal diversity in loog-pang and koji starters, respectively. The results suggest that loog-pang and koji can produce cellulase, neutral and acid proteases during fermentation. Despite the discrepancy in their microbial diversity and the enzyme activity during SSF, both starters could effectively increase protein extraction from HSDRB. A positive relationship between the SSF duration and extracted protein was observed. During SSF with loog-pang and koji after 72 h followed by 24-hour hydrolysis, 65.66 and 66.67 % protein was extracted from HSDRB, respectively. The amino acid analysis of the protein hydrolysate produced by the non-fermented and fermented methods showed no difference and had an abundance of glutamic and aspartic acids, leucine, arginine, alanine and glycine amino acids, which accounted for approx. 58 % of the total amino acids.
Loog-pang and koji (traditional Thai and Japanese fermentation starters, respectively) were found to be effective in extracting proteins from HSDRB by SSF although they are inexpensive microbial enzyme sources. Future research aimed at scaling up HSDRB protein extraction for usage in industrial applications can draw on our results.
热稳定脱脂米糠(HSDRB)是米糠油提取行业的主要副产品,也是一种营养丰富的蛋白质来源。然而,尽管米糠蛋白具有独特的营养特性,但这种富含蛋白质的副产品HSDRB却作为低价值动物饲料未得到充分利用。通过酶水解从HSDRB中提取蛋白质的研究引起了众多科学家的关注。然而,需要一种经济高效的提取方法来降低与酶使用相关的高成本。因此,我们提出了一种通过异养发酵微生物进行固态发酵(SSF)从HSDRB中提取蛋白质的替代经济且天然的方法。
评估了使用两种传统亚洲发酵剂,即鲁邦(loog - pang)和酒曲(koji)对HSDRB进行固态发酵以生产酶及其从HSDRB中提取蛋白质的功效。为此,对HSDRB进行0、12、24、48、72和96小时的发酵,随后进行24小时水解以评估提取的米糠蛋白。此外,还通过对16S rRNA和内转录间隔区进行宏基因组测序分别确定发酵剂中的微生物群落多样性,以鉴定细菌和真菌。
发酵剂中的微生物群落显示乳酸菌(LAB)占优势,如鲁邦中的[具体乳酸菌名称1]和[具体乳酸菌名称2],以及酒曲中的[具体乳酸菌名称3]和[具体乳酸菌名称4],而酵母物种[具体酵母名称1]和[具体酵母名称2]分别在鲁邦和酒曲发酵剂的真菌多样性中占主导地位。结果表明,鲁邦和酒曲在发酵过程中可产生纤维素酶、中性和酸性蛋白酶。尽管在固态发酵过程中它们的微生物多样性和酶活性存在差异,但两种发酵剂都能有效提高从HSDRB中提取蛋白质的量。观察到固态发酵时间与提取的蛋白质之间存在正相关关系。在使用鲁邦和酒曲进行72小时固态发酵后再进行24小时水解,分别从HSDRB中提取了65.66%和66.67%的蛋白质。对未发酵和发酵方法产生的蛋白质水解产物的氨基酸分析表明没有差异,并且富含谷氨酸、天冬氨酸、亮氨酸、精氨酸、丙氨酸和甘氨酸,这些氨基酸约占总氨基酸的58%。
尽管鲁邦和酒曲(分别为传统泰国和日本发酵剂)是廉价的微生物酶源,但发现它们通过固态发酵从HSDRB中提取蛋白质是有效的。旨在扩大HSDRB蛋白质提取以用于工业应用的未来研究可以借鉴我们的结果。