• Journal of Microbiology and Biotechnology
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• 제21권3호
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• pp.284-292
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• 2011

An endocellulase-free multienzyme complex was produced by a thermophilic anaerobic bacterium, Thermoanaerobacterium thermosaccharolyticum strain NOI-1, when grown on xylan. The temperature and pH optima for growth were $60^{\circ}C$ and 6.0, respectively. The bacterial cells were found to adhere to insoluble xylan and Avicel. A scanning electron microscopy analysis showed the adhesion of xylan to the cells. An endocellulase-free multienzyme complex was isolated from the crude enzyme of strain NOI-1 by affinity purification on cellulose and Sephacryl S-300 gel filtration. The molecular mass of the multienzyme complex was estimated to be about 1,200 kDa. The multienzyme complex showed one protein on native PAGE, one xylanase on a native zymogram, 21 proteins on SDS-PAGE, and 5 xylanases on a SDS zymogram. The multienzyme complex consisted of xylanase, ${\beta}$-xylosidase, ${\alpha}$-L-arabinofuranosidase, ${\beta}$-glucosidase, and cellobiohydrolase. The multienzyme complex was effective in hydrolyzing xylan and corn hulls. This is the first report of an endocellulase-free multienzyme complex produced by a thermophilic anaerobic bacterium, T. thermosaccharolyticum strain NOI-1.

• Journal of Animal Science and Technology
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• 제66권2호
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• pp.398-411
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• 2024

Upregulation of the nutritional value of feed is the major target of various studies in the livestock industry, and dietary enzyme supplementation could aid in digesting the nondegrading nutrients of grains in feed ingredients. Dried distillers' grains with solubles (DDGS) is a byproduct of the fermentation process in the beverage industry and can be used as a large supply source of fiber in feed. Therefore, we conducted an experiment with male broiler chickens to investigate the effect of various types of enzymes on DDGS and compare the efficacy of single enzyme and multienzyme complexes on growth performance and gut environments in broiler chickens. We used 420 1-day-old broiler chickens (Ross 308), and they were allotted into 4 dietary treatments with seven replications (CON, corn-soybean meal [SBM] diet; NC, DDGS supplemented diet; SE, 0.05 % of mannanase supplemented DDGS-based diet; MC, 0.10% of multienzyme complex (mannanase and xylanase, glucanase) supplemented DDGS-based diet. The dietary exogenous enzyme in the DDGS-supplemented diet could improve growth performance as much as the growth of the control group, and digestibility of dry matter, crude protein, and gross energy were significantly increased by enzyme addition in groups of chicks fed DDGS-supplementation diet. Moreover, the populations of pathogenic bacteria, coliforms, and Bacteroidetes were significantly decreased by enzyme supplementation, which might lead to improved gut mucus-secreting cells and inflammatory cytokines in the jejunum. Collectively, dietary single enzyme and multienzyme complexes could improve gut environments, including intestinal immune responses and gut microbial population, and lead to improvement of growth performance in broiler chickens.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1269-1275
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• 2006

The selection of multienzyme complex-producing bacteria under aerobic condition was conducted for improving the degradation of lignocellulosic substances. The criteria for selection were cellulase and xylanase enzyme production, the presence of cellulose-binding domains and/or xylan-binding domains in enzymes to bind to insoluble substances, the adhesion of bacterial cells to insoluble substances, and the production of multiple cellulases and xylanases in a form of a high molecular weight complex. Among the six Bacillus strains, isolated from various sources and deposited in our laboratory, Paenibacillus curdlanolyticus B-6 strain was the best producer of cellulase and xylanase enzymes, which have both cellulose-binding factors (CBFs) and xylan-binding factors (XBFs). Moreover, multiple carboxymethyl cellulases (CMCases) and xylanases were produced by the strain B-6. The zymograms analysis showed at least 9 types of xylanases and 6 types of CMCases associated in a protein band of xylanase and cellulase with high molecular weight. These cells also enabled to adhere to both avicel and insoluble xylan, which were analyzed by scanning electron microscopy. The results indicated that the strain B-6 produced the multienzyme complex, which may be cellulosome or xylanosome. Thus, P. curdlanolyticus B-6 was selected to study the role and interaction between the enzymes and their substrates and the cooperation of multiple enzymes to enhance the hydrolysis due to the complex structure for efficient cellulases and xylanases degradation of insoluble polysaccharides.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.77-83
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• 2017

Lignocellulosic biomass represents a potentially large resource to supply the world's fuel and chemical feedstocks. Enzymatic bioconversion of this substrate offers a reliable strategy for accessing this material under mild reaction conditions. Owing to the complex nature of lignocellulose, many different enzymatic activities are required to function in concert to perform efficient transformation. In nature, large multienzyme complexes are known to effectively hydrolyze lignocellulose into constituent monomeric sugars. We created artificial complexes of enzymes, called rosettazymes, in order to hydrolyze glucuronoxylan, a common lignocellulose component, into its cognate sugar ${\small{D}}$-xylose and then further convert the ${\small{D}}$-xylose into ${\small{D}}$-xylonic acid, a Department of Energy top-30 platform chemical. Four different types of enzymes (endoxylanase, ${\alpha}$-glucuronidase, ${\beta}$-xylosidase, and xylose dehydrogenase) were incorporated into the artificial complexes. We demonstrated that tethering our enzymes in a complex resulted in significantly more activity (up to 71%) than the same amount of enzymes free in solution. We also determined that varying the enzyme composition affected the level of complex-related activity enhancement as well as overall yield.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 1998년도 학술발표회
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• pp.20-20
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• 1998

In mammalian cells, nine aminoacyl-tRNA synthetase, including aspartyl-tRNA synthetase, are associated within a multienzyme complex. Human aspartyl-tRNA synthetase contains a unique N-terminal polypeptide that is thought to be responsible for the complex formation.(omitted)

• KSBB Journal
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• 제30권2호
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• pp.63-68
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• 2015

This study aimed to develop and optimize a demucilaging process of Opuntia ficus-indica var. saboten (OFI) fruit to increase its usability as functional food ingredient and food additive. Viscozyme and Novozym 33095 as multienzyme complex having a broad spectrum of carbohydrases and pectolytic enzymes, respectively, were used in enzymatic dissolution along with high hydrostatic pressure liquefaction. To optimize the liquefaction process using high hydrostatic pressure liquefying extractor, response surface methodology with 3-factor central composite design was employed with reaction factors such as temperatures (25, 32, 40, 48, and $55^{\circ}C$), pressures (20, 40, 60, 80, and 100 MPa), and times (15, 30, 45, 60, and 75 min). At optimum conditions ($25^{\circ}C$, 100 MPa, and 58.275 min) for high hydrostatic pressure liquefaction process, the processed OFI fruit juice was predicted to have viscosity at 2.917 poise, partly due to the release of free sugars such as fructose and glucose detected using HPLC-ELSA system. The results above suggests that the OFI fruit juice with decreased viscosity may be used for various manufacturing processes of food, beverage, ice cream, and cosmetics.

• 한국식품과학회지
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• 제26권5호
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• pp.484-489
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• 1994

두유 및 두부제조시 부생되는 비지를 효과적으로 이용하기 위한 방안으로 Aspergillus niger CF-34로부터 얻은 효소를 비지에 처리하여 가용화를 시도하였다. 비지가용화를 위한 본 연구에서 비지에 작용하는 복합효소액의 최적반응조건을 조사하였으며, 복합효소액 처리시 두유비지내 불용성 단백질의 회수율 및 고형분 용해도와 비지의 효소가수분해 특성을 검토하였고, 그 결과는 다음과 같다. 복합효소액의 사용량은 비지 고형분의 약 50%를 가용화하는 2.5%(% of total solid)를 최적 복합효소액의 농도로 선정하였다. 효소작용시간에 따른 비지의 단백질 회수율 및 고형분의 용해도는 효소반응후 3시간에서 각각 약 62%, 50%로 상대적으로 높은 값을 나타내었다. 비지를 alkali 처리(0.1% NaOH)로 입자구조를 변형시킴으로서 효소가수분해율이 증가되었고, 가수분해 시간도 단축됨에 따라 비지 가수분해의 효과적인 전처리방법임을 확인할 수 있었다.