A new engineered endo-inulinase with improved activity and thermostability: Application in the production of prebiotic fructo-oligosaccharides from inulin

Mao, Weian; Han, Yao-Zu; Wang, Xiaoxiang; Zhao, Xiaoxue; Chi, Zhe; Chi, Zhenimg; Liu, Guanglei

doi:10.1016/j.foodchem.2019.05.062

Cited by 22 publications

(7 citation statements)

References 36 publications

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“…The C-terminal fusion one (Eninu-IBM) with high activity, thermal stability, and inulin binding efficiency was used to catalyze inulin break down at high-temperature in a 10-L bioreactor. Results obtained high purity FOS (91.4%) with FOS titer, productivity, and yield of 717.3 g/L, 358.6 g/L/h and 0.912 gFOS/g inulin, respectively ( Mao et al, 2019 ). Risso et al (2012) investigated the FOS production by carrier-supported inulinase from K. marxianus NRRL Y-7571 in aqueous and aqueous-organic systems using sucrose as a substrate.…”

Section: Applications In Biotechnological Productsmentioning

confidence: 99%

Bioprospecting Kluyveromyces marxianus as a Robust Host for Industrial Biotechnology

Bilal

et al. 2022

Front. Bioeng. Biotechnol.

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Kluyveromyces marxianus is an emerging non-conventional food-grade yeast that is generally isolated from diverse habitats, like kefir grain, fermented dairy products, sugar industry sewage, plants, and sisal leaves. A unique set of beneficial traits, such as fastest growth, thermotolerance, and broad substrate spectrum (i.e., hemi-cellulose hydrolysates, xylose, l-arabinose, d-mannose, galactose, maltose, sugar syrup molasses, cellobiose, and dairy industry) makes this yeast a particularly attractive host for applications in a variety of food and biotechnology industries. In contrast to Saccharomyces cerevisiae, most of the K. marxianus strains are apparently Crabtree-negative or having aerobic-respiring characteristics, and unlikely to endure aerobic alcoholic fermentation. This is a desirable phenotype for the large-scale biosynthesis of products associated with biomass formation because the formation of ethanol as an undesirable byproduct can be evaded under aerobic conditions. Herein, we discuss the current insight into the potential applications of K. marxianus as a robust yeast cell factory to produce various industrially pertinent enzymes, bioethanol, cell proteins, probiotic, fructose, and fructo-oligosaccharides, and vaccines, with excellent natural features. Moreover, the biotechnological improvement and development of new biotechnological tools, particularly CRISPR–Cas9-assisted precise genome editing in K. marxianus are delineated. Lastly, the ongoing challenges, concluding remarks, and future prospects for expanding the scope of K. marxianus utilization in modern biotechnology, food, feed, and pharmaceutical industries are also thoroughly vetted. In conclusion, it is critical to apprehend knowledge gaps around genes, metabolic pathways, key enzymes, and regulation for gaining a complete insight into the mechanism for producing relevant metabolites by K. marxianus.

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Section: Applications In Biotechnological Productsmentioning

confidence: 99%

Bioprospecting Kluyveromyces marxianus as a Robust Host for Industrial Biotechnology

Bilal

et al. 2022

Front. Bioeng. Biotechnol.

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“…Endo-inulinase (EC3.2.1.7) is the enzyme used for FOS production from inulin. This endo-inulinase enzyme hydrolyze the 2,1-β-D-glycosidic bonds in inulin chemical structure for the production of FOS as end products [5].…”

Section: Endo-inulinase (Ec3217)mentioning

confidence: 99%

Technological Aspects of Fructo-Oligosaccharides (FOS), Production Processes, Physiological Properties, Applications and Health Benefits

Ibrahim¹

2021

J Food Chem Nanotechnol

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Fructo-oligosaccharides (FOS) has two other names oligofructose and oligofructan are naturally presents in fruits and vegetables but are mainly produced enzymatically from sucrose or inulin as enzymes substrates. Its chemical structures are short chain of fructose polymer with terminal glucose unit comparing to inulin that has similar chemical structure but at higher degree of fructose polymerization (DP). Inulin is present at high level in Chicory roots and Jerusalem artichoke. The enzyme fructosyl-transferase or the enzyme ß-fructo-furanosidase are used for the production of FOS from sucrose as substrate, while the enzyme endoinulinase is used for the production of FOS from inulin as a substrate. Fructooligosaccharides (FOS) are used in food applications, dietary supplements and nutrition bars as low-calorie sweetener for diabetes in replacement to sucrose, also used in pharmaceutical drugs and cosmetics products formulation. FOS for health benefits is used in dietary fiber and as prebiotics for constipation and to enhance the growth of beneficial bacteria in the colon respectively. Other health benefits for FOS are increase mineral absorption in digesting system, and decrease, serum cholesterol/triglycerides. FOS are General Recognized as Safe (GRAS) in United States, and also approved for use by other international regulatory agencies in food products, infant formulas, pharmaceutical and in other applications.

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“…Inulin, composed of β‐2,1‐linked D‐fructosyl residues, is a reserve carbohydrate present in a range of plant species such as Jerusalem artichoke, chicory, dahlia, and yacon (Singh et al, 2019). Recently, inulin and inulin‐containing materials have attracted considerable research attention as nongrain biomass for the production of various microbial products, such as high fructose syrup, fructooligosaccharides, bioethanol, organic acids, single‐cell oils, and single‐cell proteins (Liu et al, 2010, 2013, 2014; Mao et al, 2019; Singh et al, 2019). Inulinase is a fructofuranosyl hydrolase that targets the β‐2,1‐linkage of inulin and can be produced by a wide range of microorganisms (Kango & Jain, 2011; Liu et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A novel transcriptional activation mechanism of inulinase gene in Kluyveromyces marxianus involving a glycolysis regulator KmGcr1p with unique and functional Q‐rich repeats

Wang

Chi

et al. 2022

Molecular Microbiology

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Kluyveromyces marxianus is the most suitable fungus for inulinase industrial production. However, the underlying transcriptional activation mechanism of the inulinase gene (INU1) is hitherto unclear. Here, we undertook genetic and biochemical analyses to elucidate that a glycolysis regulator KmGcr1p with unique Q‐rich repeats is the key transcriptional activator of INU1. We determined that INU1 and glycolytic genes share similar transcriptional activation patterns and that inulinase activity is induced by fermentable carbon sources including the hydrolysis products of inulin (fructose and glucose), which suggests a novel model of product feedback activation. Furthermore, all four CT‐boxes in the INU1 promoter are important for KmGcr1p DNA‐binding in vitro, but the most downstream CT‐box 1 primarily confers upstream activating sequence activity in vivo. More intriguingly, the use of artificial and natural GCR1 mutants suggests that the Q‐rich repeats act as a functional module to maintain KmGcr1p transcriptional activity by contributing to its solubility and DNA‐binding affinity. Altogether, this study uncovers a novel transcriptional activation mechanism for the inulinase gene, that is different from the previous understanding for filamentous fungi, but might have universal significance among inulinase‐producing yeasts, thereby leading to a better understanding of the regulation mechanism of yeast inulinase genes.

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A new engineered endo-inulinase with improved activity and thermostability: Application in the production of prebiotic fructo-oligosaccharides from inulin

Cited by 22 publications

References 36 publications

Bioprospecting Kluyveromyces marxianus as a Robust Host for Industrial Biotechnology

Bioprospecting Kluyveromyces marxianus as a Robust Host for Industrial Biotechnology

Technological Aspects of Fructo-Oligosaccharides (FOS), Production Processes, Physiological Properties, Applications and Health Benefits

A novel transcriptional activation mechanism of inulinase gene in Kluyveromyces marxianus involving a glycolysis regulator KmGcr1p with unique and functional Q‐rich repeats

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