Mafa Mpho S, Malgas Samkelo, Pletschke Brett I
Enzyme Science Programme (ESP), Department of Biochemistry and Microbiology, Rhodes University, Grahamstown, 6140, South Africa.
Department of Plant Sciences, University of the Free State, P.O. Box 339, Bloemfontein, 9300, South Africa.
AMB Express. 2021 Jan 19;11(1):21. doi: 10.1186/s13568-021-01180-1.
Cereal feedstocks have high arabinoxylan content as their main hemicellulose, which is linked to lignin by hydroxycinnamic acids such as ferulic acid. The ferulic acid is linked to arabinoxylan by ester bonds, and generally, the high substitution of ferulic acid leads to a loss of activity of xylanases targeting the arabinoxylan. In the current study, a feruloyl esterase (FAE-1) from a termite hindgut bacteria was functionally characterised and used in synergy with xylanases during xylan hydrolysis. The FAE-1 displayed temperature and pH optima of 60 ℃ and 7.0, respectively. FAE-1 did not release reducing sugars from beechwood xylan (BWX), wheat arabinoxylan (WAX) and oat spelt xylan (OX), however, displayed high activity of 164.74 U/mg protein on p-nitrophenyl-acetate (pNPA). In contrast, the GH10 xylanases; Xyn10 and XT6, and a GH11 xylanase, Xyn2A, showed more than two-fold increased activity on xylan substrates with low sidechain substitutions; BWX and OX, compared to the highly branched substrate, WAX. Interestingly, the FAE-1 and GH10 xylanases (Xyn10D and XT6) displayed a degree of synergy (DS) that was higher than 1 in all enzyme loading combinations during WAX hydrolysis. The 75%XT6:25%FAE-1 synergistic enzyme combination increased the release of reducing sugars by 1.34-fold from WAX compared to the control, while 25%Xyn10D:75%FAE-1 synergistic combination released about 2.1-fold of reducing sugars from WAX compared to controls. These findings suggest that FAE-1 can be used in concert with xylanases, particularly those from GH10, to efficiently degrade arabinoxylans contained in cereal feedstocks for various industrial settings such as in animal feeds and baking.
谷物原料中的主要半纤维素是阿拉伯木聚糖含量很高,它通过阿魏酸等羟基肉桂酸与木质素相连。阿魏酸通过酯键与阿拉伯木聚糖相连,一般来说,阿魏酸的高取代会导致靶向阿拉伯木聚糖的木聚糖酶活性丧失。在本研究中,对一种来自白蚁后肠细菌的阿魏酸酯酶(FAE-1)进行了功能表征,并在木聚糖水解过程中与木聚糖酶协同使用。FAE-1的温度和pH最佳值分别为60℃和7.0。FAE-1不能从山毛榉木聚糖(BWX)、小麦阿拉伯木聚糖(WAX)和燕麦spel木聚糖(OX)中释放还原糖,然而,它对对硝基苯乙酸(pNPA)显示出164.74 U/mg蛋白质的高活性。相比之下,GH10木聚糖酶Xyn10和XT6以及GH11木聚糖酶Xyn2A在低侧链取代的木聚糖底物(BWX和OX)上的活性比高度分支的底物WAX增加了两倍以上。有趣的是,在WAX水解过程中,FAE-1和GH10木聚糖酶(Xyn10D和XT6)在所有酶负载组合中都表现出协同度(DS)高于1。与对照相比,75%XT6:25%FAE-1协同酶组合使WAX中还原糖的释放增加了1.34倍,而25%Xyn10D:75%FAE-1协同组合使WAX中还原糖的释放量比对照增加了约2.1倍。这些发现表明,FAE-1可以与木聚糖酶协同使用,特别是那些来自GH10的木聚糖酶,以有效降解谷物原料中含有的阿拉伯木聚糖,用于各种工业环境,如动物饲料和烘焙。