scholarly journals Statistical-Based Bioprocess Optimization of Amylase Production from Halophilic Bacillus sp. H7.

Author(s):  
J.N. Bandal ◽  
V.A. Tile ◽  
R. Z. Sayyed ◽  
H.P. Jadhav ◽  
N. I. Wan Azelee ◽  
...  

Using the above results from RMS analysis the optimum values were predicted for the independent significant variables (Figure 3) the optimized levels of these variables in combination with other media variables the maximum production was predicted to be 199.90 U/mL. The predicted data were validated through confirmatory experiments performed in triplicates. A 1.29-fold increase in amylase activity against un-optimized (OVAT) medium was achieved in the present study authenticating the efficacy of RSM in process optimization (Figure 4). 2.6 Model validation and scale-up at laboratory scale (5L) bioreactor Once the parameters were standardized in the shake-flasks culture, the experiment was scaled-up to a laboratory-scale bioreactor (5 L). The yield of amylase increased by 1.01 fold (205.69 U/mL), it could be possible because the enzyme production in a bioreactor is higher than in shake-flasks culture as the various critical variable factors such as the dissolved oxygen (DO) and the pH can be optimally controlled at the desired levels [22].

2017 ◽  
Vol 7 (3) ◽  
pp. 7 ◽  
Author(s):  
Rachna Singh ◽  
Deepak Chand Sharma ◽  
Mahendra Kumar Gupta

The aim of this work was to optimize the process parameters through the statistical approach for the production of alpha amylase by Bacillus sp. in submerged fermentation (SMF). Initially critical physical and chemical process parameters influencing the enzyme production were identified by Plackett-Burman method (eleven variables, seven nutritional, two physical and two dummies) were analyzed. Then optimum levels of most influencing parameters affecting amylase production were obtained by CCD and results were analyzed by standard analysis of variance (ANOVA). The effect of interaction of physiochemical parameters on the amylase production (z axis) was studied by plotting three dimensional response surface curves against any two independent variables. A high similarity was observed between the predicted and experimental results, which reflected the accuracy and applicability of RSM to optimize the process for enzyme production. As a result of RSM the optimum values for starch concentration -0.65%, (NH4)2SO4 -0.55% and pH- 8.33. As a result of media optimization, a titre of 1, 22,000 U/L-1 was achieved. A total of 2.4 fold increase in enzyme activity was observed.


Author(s):  
Ewa Kochan ◽  
Sylwia Caban ◽  
Grażyna Szymańska ◽  
Piotr Szymczyk ◽  
Anna Lipert ◽  
...  

<p>Plant suspension cultures are described as a source for the acquisition of medicinal secondary metabolites which in the future may become an alternative to traditional raw materials. This study demonstrates that the cell cultures of one of the ginseng species – Panax quinquefolium L. synthesize ginsenosides, which are triterpene saponins having a multidirectional pharmacological effects. Tested suspension cultures were run on a small scale in the shake flasksand in scale up of the process in a 10-liter stirred tank. In the shake flasks,the highest biomass yield (2.28 gl-1 for dry and 33.99 gl-1 for fresh weight) was reached on day 30 of culture, and the highest content of saponins (2.66 mg g -1 dw) was determined on day 28 of culture. In the bioreactor, nearly 2.67 and 3-fold increase of respectively dry and fresh biomass was recorded in relation to the inoculum. Large-scale cultures synthesized protopanaxatriol derivatives such as Rg1 and Re ginsenosides, however, no saponins belonging to the protopanaxadiol derivatives were reported.</p>


2021 ◽  
Author(s):  
Most Sheauly Khatun ◽  
Morteza Hassanpour ◽  
Mark Harrison ◽  
Robert Speight ◽  
Ian O'Hara ◽  
...  

Abstract In this study, sugarcane molasses was used to produce transfructosylating enzymes by A. pullulans FRR 5284. It was found that NaNO3 was a better nitrogen source than yeast extract while exogeneous phosphorous was not needed. Adding only 4.4 g/L NaNO3 into the molasses medium containing 100 g/L sugars led to the highest total transfructosylating activity of 123.8 U/mL. Scale-up of the enzyme production process from shake flasks to 1 L reactor improved the enzyme activity and productivity to 171.7 U/mL and 3.58 U/mL/h, 39% and 108% higher than the corresponding activity and productivity from shake flasks, respectively. FOS production from 500 g/L sucrose led to the highest yields of ~ 61% using intracellular, extracellular, and total enzymes from shake flasks and the reactor. Enzymes from different sources led to very different FOS profiles, indicating that FOS profiles can be controlled by adjusting intracellular and extracellular enzyme ratios to adjust prebiotic activity.


1970 ◽  
Vol 24 (2) ◽  
pp. 129-132 ◽  
Author(s):  
Arifa Nusrat ◽  
Sabita Rezwanan Rahman

In the present study three mesophilic Bacillus isolates were analyzed for their α-amylase activity in shakeflask cultures. The organisms were capable to produce hydrolysis zone around their colonies on starch agar medium. The effect of various fermentation conditions on α-amylase production was investigated, and in every case it was found that B. subtilis was the best producer of the enzyme, which was followed by the newly isolated Bacillus sp. and B. amyloliquefaciens. The synthesis of extracellular α-amylase by the bacteria was repressed by the presence of readily metabolizable carbon source like glucose in the culture medium. Maximum α-amylase activity by the Bacillus isolates was obtained at 37°C with an initial medium pH 7.0 under agitation at 160-180 rpm for 72 h of growth. Keywords: Bacillus species, α-Amylase, Enzyme production, Shake-flask cultureDOI: http://dx.doi.org/10.3329/bjm.v24i2.1257 Bangladesh J Microbiol, Volume 24, Number 2, December 2007, pp 129-132


2019 ◽  
Vol 17 ◽  
Author(s):  
Farzane Kargar ◽  
Mojtaba Mortazavi ◽  
Mahmood Maleki ◽  
Masoud Torkzadeh Mahani ◽  
Younes Ghasemi ◽  
...  

Aims: The purpose of this study was to screen the bacteria producing cellulase enzymes and their bioinformatics studies. Background: Cellulose is a long-chain polymer of glucose that hydrolyzes by cellulases to glucose molecules. In order to design the new biotechnological applications, some strategies have been used as increasing the efficiency of enzyme production, generating cost-effective enzymes, producing stable enzymes and identification of new strains. Objective: On the other hand, some bacteria special features have made them suitable candidates for the identification of the new source of enzymes. In this regard, some native strains of bacteria were screened. Method: These bacteria were grown on a culture containing the liquid M9 media containing CMC to ensure the synthesis of cellulase. The formation of a clear area in the culture medium indicated decomposition of cellulose. In the following, the DNA of these bacteria were extracted and their 16S rDNA genes were amplified. Result: The results show that nine samples were able to synthesize cellulase. In following, these strains were identified using 16S rDNA. The results show that these screened bacteria belonged to the Bacillus sp., Alcaligenes sp., Alcaligenes sp., and Enterobacter sp.conclusionThe enzyme activity analysis shows that the Bacillus toyonensis, Bacillus sp. strain XA15-411 Bacillus cereus have produced the maximum yield of cellulases. However, these amounts of enzyme production in these samples are not proportional to their growth rate. As the bacterial growth chart within 4 consecutive days shows that the Alcaligenes sp. Bacillus cereus, Bacillus toyonensis, Bacillus sp. strain XA15-411 have a maximum growth rate. The study of the phylogenetic tree also shows that Bacillus species are more abundant in the production of cellulase enzyme. These bioinformatics analyses show that the Bacillus species have different evolutionary relationships and evolved in different evolutionary time. Other: However, for maximum cellulase production by this bacteria, some information as optimum temperature, optimum pH, carbon and nitrogen sources are needed for the ideal formulation of media composition. The cellulase production is closely controlled in microorganisms and the cellulase yields appear to depend on a variety of factors. However, the further studies are needed for cloning, purification and application of these new microbial cellulases in the different commercial fields as in food, detergent, and pharmaceutical, paper, textile industries and also various chemical industries. However, these novel enzymes can be further engineered through rational design or using random mutagenesis techniques.


2001 ◽  
Vol 56 (11-12) ◽  
pp. 1022-1028 ◽  
Author(s):  
Kristina Uzunova ◽  
Anna Vassileva ◽  
Margarita Kambourova ◽  
Viara Ivanova ◽  
Dimitrina Spasova ◽  
...  

Abstract Enzyme production of newly isolated thermophilic inulin-degrading Bacillus sp. 11 strain was studied by batch cultivation in a fermentor. The achieved inulinase and invertase activi­ ties after a short growth time (4.25 h) were similar or higher compared to those reported for other mesophilic aerobic or anaerobic thermophilic bacterial producers and yeasts. The investigated enzyme belonged to the exo-type inulinases and splitted-off inulin, sucrose and raffinose. It could be used at temperatures above 65 °C and pH range 5.5-7.5. The obtained crude enzyme preparation possessed high thermostability. The residual inulinase and inver­ tase activities were 92-98% after pretreatment at 65 °C for 60 min in the presence of substrate inulin.


2003 ◽  
Vol 46 (3) ◽  
pp. 455-459 ◽  
Author(s):  
Georgina L. Michelena ◽  
Aidín Martínez ◽  
Antonio Bell ◽  
Emilia Carrera ◽  
Roxana Valencia

Fed batch fermentation was carried out for the dextransucrase enzyme production from Leuconostoc mesenteroides and the production was scale-up using oxygen transfer criteriuom. It was found that in 5 L vessel fermentation capacity, the best agitation speed was 225 min-1 and aeration rate was 0.15 vvm, obtaining dextransucrase activity of 127 DSU/mL.. The maximum enzyme production velocity coincide with the maximum growth velocity between 6 and 7 h of fermentation, which confirmed that dextransucrase production was associated with microbial growth. High enzyme yields were achieved during scale up based on oxygen transfer rate.


2016 ◽  
Vol 3 (2) ◽  
pp. 134-146
Author(s):  
A. A. Fatyanova ◽  
A. S. Gusak ◽  
P. E. Prokhorova ◽  
O. A. Trofimova
Keyword(s):  
Scale Up ◽  

2018 ◽  
Vol 25 (03) ◽  
pp. 80-89
Author(s):  
Naranchimeg B ◽  
Altantsetseg Kh ◽  
Urantulkhuur B

Amylase is one of the most widely used enzymes in many industrial sectors (starch decomposition, bakery, fermentation, biofuel, detergent, paper, textile, etc.), thus isolating pure cultures of amylase producing microorganisms from natural sources and improving their activity is important in biotechnology. Enzyme preparations with high activity can be obtained only by improved synthesis of biologically active substances of microorganisms by mutagenesis. In the present investigation was enhanced the amylase productivity of some Bacillus sp. (assigned as 1,2,3) isolated from soil sample by substrate induction and mutagenesis. 3 isolates are subcultured in the medium with starch (10 mg/ml) as only carbon source, to improve amylase production. Enzyme activity of parental strains increased 50-58% by substrate induction. The highest productive strain (Bacillus sp. 2) screened and selected. Then it was subjected to 4 period mutagenesis using UV irradiation and еthidium bromide. Amylase activity of Bacillus sp. 2 increased after first period of mutagenesis to:0.305,  second:0.514, third:0.579 and fourth:0.592 U/ml. In the result our experiment, amylase activity of parental strain increased from 0.138 to 0.592 U/ml, which means 4.3 times more enzyme. Амилаза нийлэгжүүлэгч бактерийн мутант омог гарган авсан дүн Хураангуй: Амилаза нь үйлдвэрлэлийн олон салбарт (цардуул задлах, талх нарийн боов, исгэлт, биотүлш, угаалгын нунтаг, цаас, нэхмэл гэх мэт) өргөнөөр хэрэглэгддэг фермент тул түүнийг нийлэгжүүлэгч бичил биетний өсгөврийг байгалийн эх үүсвэрээс ялган авч, идэвхийг нь сайжруулах явдал биотехнологийн салбарт чухал ач холбогдолтой юм. Мутагенезийн аргаар бичил биетний биологийн идэвхтэй бодисын нийлэгжилтийг сайжруулснаар ферментийн бэлдмэл гарган авах боломжтой болно. Судалгаагаар Монгол орны биосферээс ялган авсан цардуул задлах идэвхтэй бактерийн цэвэр өсгөврийн амилаза ферментийн идэвхийг субстратаар өдөөх болон мутагенезийн аргаар сайжруулахыг зорив. Хөрснөөс ялган авсан цардуул задлах идэвхтэй цэвэр өсгөврүүдийг (Bacillus sp. 1, 2, 3) нүүрс-усны эх үүсвэрээр зөвхөн цардуул агуулсан (10 г/л) тэжээлт орчинд өсгөвөрлөх замаар амилаза ферментийн идэвхийг нэмэгдүүлэх туршилт хийж үр дүнг үндэслэн хамгийн идэвхтэй нэг өсгөврийг сонгон шалгаруулж, хэт ягаан туяа, этидиум бромид, хэт ягаан туяа, этидиум бромид гэсэн дарааллаар зориудын мутагенезид 4 үе шаттайгаар оруулав. Субстратаар өдөөхөд бактерийн амилаза ферментийн идэвх анхдагч өсгөврийнхөөс 50-58 хувиар нэмэгдсэн. Bacillus sp. 2 өсгөврийг сонгон шалгаруулж, мутагенезид оруулахад амилаза ферментийн идэвх нь I шатны мутагенезээр:0,305, II-оор:0,514, III-аар:0,579, IV-өөр:0,592 н/мл болж нэмэгдэв. Бидний судалгааны үр дүнд байгалийн анхдагч өсгөврийн (Bacillus sp. 2) амилаза ферментийн идэвх 0,138-аас 0,592 н/мл хүртэл буюу 4,3 дахин нэмэгдсэн байна. Түлхүүр үг: амилаза, Bacillus sp., субстратын өдөөлт, UV-мутагенез, EtBr- мутангенез


Sign in / Sign up

Export Citation Format

Share Document