Overexpression of hemA and hemL in Bacillus subtilis Promotes Overexpression of 5-aminolevulinic Acid 

2020 ◽  
Author(s):  
Jinni Liu ◽  
Zhaowei Ye ◽  
Haigang Wu ◽  
Jicheng Liu ◽  
Yuesheng Gong
Keyword(s):  
Bacillus Subtilis ◽  
Acid Content ◽  
Soluble Protein ◽  
Aminolevulinic Acid ◽  
Culture Media ◽  
Total Soluble Protein ◽  
Target Proteins

Bacillus subtilis strains were constructed that overexpressed the glutamyl t-RNA reductase (HemA) and glutamyl t-RNA synthase (HemL). The target proteins accounted for 11 and 13% of the total soluble protein in maltose induced extracts, respectively. The culture media from these strains was red with wavelength maxima near 400 nm indicating elevated levels of porphyrins. The 5-aminolevulinic acid content for these strains reached 42.57 and 39.78 mg/L after 24 h of induced growth and 65.23 and 68.45 mg/L, respectively, after 60 h. Therefore, hemA and hemL overexpression enhanced the biosynthesis of 5-aminolevulinic acid and these strains can be used for its purification.

Genetic and Biochemical Characterization of Electron Beam and Gamma Ray Induced Mutants for Superior Battering Quality Trait and MYMV in Urdbean [Vigna mungo (L.)]

2021 ◽  
Author(s):  
C. Vanniarajan ◽  
R. Chandirakala ◽  
J. Souframanien
Keyword(s):  
Phytic Acid ◽  
Acid Content ◽  
Soluble Protein ◽  
Biochemical Characterization ◽  
Total Phenol ◽  
Total Soluble Protein ◽  
Yellow Mosaic Virus ◽  
Phytic Acid Content ◽  
Volume Analysis ◽  
Mutant Lines

Background: A systematic investigation of the induced mutagenesis in Urdbean was attempted with cultivars viz., MDU 1 and VBN (Bg) 4 to generate superior battering quality varieties. Induced mutation is proven as an applicable breeding method to build variation in plant materials. In this study, 40 Urdbean genotypes including thirty eight mutants of two promising Urdbean varieties namely MDU1 and VBN (Bg) 4 and two parental genotypes were involved for quality analysis. One of the most dangerous diseases in legumes is Mungbean Yellow Mosaic Virus (MYMV). In legume, this virus causes enormous losses in the areas of Urdbean cultivation. The prevailing varieties are partially resistant to this virus. Hence, the paramount approach to control this disease is breeding for resistance or tolerance to this disease. There is a necessity of searching the sources of disease resistance. In this study, screening of the mutants was conducted to make out the source of resistance to this disease.Methods: The nutritional characters viz., albumin, globulin and total soluble protein along with biochemical analysis of battering quality through Arabinose estimation were analyzed to find out the quality characters of the mutants. Batter volume analysis from idli batter was conducted conducted. Scoring of disease severity was done for all the plants on 50th day on the basis of visual scoring. The disease was scored on a 1-9 arbitrary scale. For biochemical characterization for this disease estimation of total phenol content, total sugars and phytic acid was executed.Result: Based on the general performance of albumin content, globulin content, total soluble protein, arabinose content, 100 seed weight and seed yield per plant, the mutant lines viz., ACM-014-021, ACM-015-015, ACM-15-023, ACM-015-013, ACM-015-003, ACM-015-030, ACM-014-006, ACM-014-007 were recognized as the best. Hence these mutant lines could be exploited directly in the breeding programme to develop superior battering quality Urdbean varieties. Based on over all mean performance of important batter volume characters viz., initial batter volume analysis, final batter volume, improved batter volume, arabinose content and single plant yield the mutant lines ACM-15-015, ACM-015-030, ACM-015-003, ACM-015-023 were identified to be superior in terms of yield, arabinose content and batter volume. The mutant lines viz., ACM-014-021, ACM-015-025, ACM-014-006, ACM-015-022, ACM -015-023, ACM-014-019, ACM-014-007, ACM-015-017, ACM-014- 003 and VBN (Bg) 4 were recognized as resistant to MYMV. These lines had higher total phenol, phytic acid content and less total sugar content in the leaves when compared with the highly susceptible mutant lines. The present study indicated that phytic acid content recorded highly significant positive correlation with total phenol. Based on the per se performance for all the characters studied viz., seed batter analysis, seed volume analysis and MYMV scoring, the mutant lines viz., ACM-014-021, ACM-015-23, ACM-015-030, were judged as the best and could be exploited directly or in breeding experiments for improving yield along with batter quality.

Tissue culture and genetic analysis of somaclonal variations of Solanum melongena L. cv. Nirrala

2014 ◽  
Vol 9 (12) ◽  
pp. 1182-1195
Author(s):  
Samar Naseer ◽  
Tariq Mahmood
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Soluble Protein ◽  
Solanum Melongena ◽  
Total Soluble Protein ◽  
Naphthalene Acetic Acid ◽  
Fresh Tissue ◽  
Somaclonal Variations ◽  
Random Amplification ◽  
Plant Grown

AbstractThe present study was designed to analyze genetically somaclonal variants using biochemical and molecular markers. Efficient tissue culture protocol for Solanum melongena L. cv. Nirrala was developed. Maximum callus induction (100%) was observed for Murashige and Skoog (MS) media supplemented with 2.0 mg L−1 naphthalene acetic acid +0.5 mg L−1 6-benzylaminopurine; and nodal explants gave best callusing response (88.8%) as compared to internodes (88.3%) and leaves (87.7%). The best shooting was induced on nodal and internodal callus in the presence of 2.0 mg L−1 6-benzylaminopurine. Total soluble protein content of callus and regenerated variant plants was estimated for biochemical analysis, and largest amount of soluble protein was found in callus (6.54 mg g−1 fresh tissue) followed by variant plant grown on 2.0 mg L−1 6-benzylaminopurine (5.96 mg g−1 fresh tissue). Random amplification of polymorphic DNA technique was done with five decamer primers (OPC1-OPC5) and maximum polymorphism was detected by OPC 2 (26.99%) among all samples, whereas nodal callus on media containing 1.0 mg L−1 naphthalene acetic acid +1.0 mg L−1 6-benzylaminopurine showed highest polymorphism producing 22 bands, out of which 8 bands were polymorphic. The study shows that this marker system can provide better evaluation of genetic variation induced by tissue culture.

scholarly journals Biochemical changes in cultured foetal rat liver explants

1975 ◽  
Vol 150 (2) ◽  
pp. 269-273 ◽  
Author(s):  
P C MacDonnel ◽  
E Ryder ◽  
J A Delvalle ◽  
O Greengard
Keyword(s):  
Soluble Protein ◽  
Cell Damage ◽  
Tyrosine Aminotransferase ◽  
Culture Conditions ◽  
Total Soluble Protein ◽  
Mitochondrial Enzymes ◽  
Stage Of Development ◽  
Foetal Rat ◽  
Total Soluble Protein Content ◽  
Do So

Liver explants from 20-day-old foetuses cultured for 48h in the absence of serum released 70% of their total soluble protein content into the medium. In the presence of serum this loss still amounted to 60%. The concentration of total particulate protein remained unchanged but there was some translocation of mitochondrial enzymes to the cytosol, and enzymes expected to increase during this stage of development failed to do so. The addition of cortisol plus glucagon (to serum-containing media) did not decrease the loss of total soluble protein from the explants but induced considerable tyrosine aminotransferase activity which was not released into the medium. The observations suggest that under the usual culture conditions a minority of the cells retain their functional integrity. The extent of deterioration, not reflected in histologically visible necrosis or cell damage, can be conveniently monitored by the malate dehydrogenase activity released to the medium.

scholarly journals Extensive destruction of newly synthesized casein in mammary explants in organ culture

1982 ◽  
Vol 202 (1) ◽  
pp. 133-138 ◽  
Author(s):  
Huthama Razooki Hasan ◽  
David A. White ◽  
R. John Mayer
Keyword(s):  
Incubation Period ◽  
Soluble Protein ◽  
Milk Protein ◽  
Fatty Acid Synthetase ◽  
Total Soluble Protein ◽  
Signal Peptides ◽  
Dual Isotope ◽  
Intracellular Degradation ◽  
Hydrolysis Of ◽  
Specific Particle

1. Explants of mammary glands of mid-pregnant rabbits that had been cultured for 18h in the presence of insulin, prolactin and cortisol were incubated at 37°C for 2h in Medium 199 containing l-[4,5-3H]leucine. After a wash procedure at 4°C, explants were re-incubated at 37°C in fresh medium and the radioactivity of casein polypeptides isolated by isoelectric focusing (at pH 4.6) was followed with time. Casein radioactivity rose during the first hour of re-incubation, but fell markedly during the subsequent hour. 2. Loss of radioactivity represented casein degradation, since less than 10% of newly synthesized casein was found in the incubation medium. 3. Such a loss of radioactivity was not due solely to hydrolysis of signal peptides, since similar results were obtained when l-[5-3H]proline, which is not part of casein signal peptides, was the radiolabelled precursor. 4. A dual-isotope experiment using l-[U-14C]proline and N-[3H]acetyl-d-mannosamine gave similar profiles of radioactivity loss from isoelectrically focused casein, indicating that degradation of mature casein was occurring. 5. Analysis of total pellet and particle-free-supernatant fractions prepared by centrifugation of explant homogenates at 115000gav. for 1h did not show loss of radioactivity on re-incubation. Total pellet-protein radioactivity remained constant, whereas total soluble-protein radioactivity increased during the 2h re-incubation period. 6. Radioactivity in a specific particle-free-supernatant polypeptide, the subunit of fatty acid synthetase, mimicked that of the total soluble protein. 7. Addition of cycloheximide (20μg/ml) during the re-incubation period completely blocked the incorporation of radioactivity from l-[5-3H]proline into casein and the subsequent fall, indicating that observations were being made on newly synthesized casein. 8. Addition of chloroquine (50μm) did not prevent the increase in radioactivity from l-[5-3H]proline into casein during the first hour of re-incubation, but did prevent the loss of radioactivity in the second hour. 9. The intracellular degradation of a newly synthesized milk protein is discussed in relation to the known intracellular degradation of other secretory polypeptides.

scholarly journals Perubahan Komposisi Kimiawi Produk Yogurt dengan Penambahan Kalsium Karbonat pada Kultur Starter Campuran

2017 ◽  
Vol 17 (1) ◽  
pp. 5
Author(s):  
Agus Safari ◽  
Sarah Fahma Ghina ◽  
Sadiah Djajasoepena ◽  
O. Suprijana ' ◽  
Ida Indrawati ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Calcium Carbonate ◽  
Chemical Composition ◽  
Lactic Acid Bacteria ◽  
Acid Content ◽  
Soluble Protein ◽  
Starter Culture ◽  
Lactobacillus Bulgaricus ◽  
Bacteria Culture

Mixed lactic acid bacteria culture is commonly used in yogurt production. In the present study, two lactic acid bacteria (Lactobacillus bulgaricus and Streptococcus thermophillus) was used as starter culture. Calcium carbonate was added to the starter culture to increase the quality of mixed starter culture of L. bulgaricus and S. thermophillus with ratio of 4:1. The present study was directed to investigate the chemical composition of mixed starter culture with and without calcium carbonat addition. Furthermore, the effect of each starter culture on yogurt product chemical composition was also examined. The pH, lactose, soluble protein and acid content was determined as chemical composition parameters. For starter culture without calcium carbonate addition, the yogurt has pH, lactose, soluble protein and acid content of 4.18–4.39, 4.18–4.39% w/v, 2.88–4.36% w/v and 0.82–0.99% w/v, respectively. While for starter culture with calcium carbonate addition, the yogurt product has pH, lactose, soluble protein and acid content of 4.26–4.37, 1.47–1.75% b/v, 3.42–4.95% w/v and 0.86–1.11% w/v, respectively. Addition of 0.05% w/v calcium carbonate to mixed starter culture gave effect on lactose consumption, where it still can convert lactose to lactic acid up to 45 days of storage. Furthermore, the yogurt product made with starter culture with calcium carbonate addition has higher soluble protein content compared to yogurt made with starter culture without calcium carbonate addition

scholarly journals Bioutilization of Chicken Feather Wastes by Newly Isolated Keratinolytic Bacteria Into Protein Hydrolysates With Improved Functionalities

2021 ◽  
Author(s):  
Saugat Prajapati ◽  
Sushil Koirala ◽  
Anil Kumar Anal
Keyword(s):  
Amino Acid ◽  
Soluble Protein ◽  
Environmental Remediation ◽  
Amino Acid Profile ◽  
Total Soluble Protein ◽  
Feather Degradation ◽  
Biochemical Tests ◽  
Box Behnken Design ◽  
Degrading Bacteria ◽  
Chicken Farm

Abstract In this study, a novel feather-degrading bacteria B. amyloliquefaciens KB1 was isolated from chicken farm bed (CFB), identified by morphological, physico-biochemical tests followed by 16s rDNA analysis. Among observed isolates, bacterial isolate (KB1) showed the highest degree of feather degradation (74.78 ± 2.94 %) and total soluble protein (205 ± 0.03 mg/ g). Using the same species of bacteria, the optimum fermentation condition was found at 40 oC, pH 9, and 1 % (w/v) feather concentration that produced 260 mg/ g of soluble protein and 86.16 % feather degradation using response surface methodology in a Box-Behnken design space. The obtained hydrolysates exhibited bioactive properties. The amino acid profile showed the increase in concentration of essential amino acid compared with feather meal broth. The selection of safe screening source of this new bacteria in CFB produced hydrolysates with enhanced bioactivity applicable for food, feed, and cosmetic applications along with environmental remediation.

scholarly journals Age-Related Changes of Some Chemical Components in the Leaves of Oriental Beech (Fagus orientalis Lipsky.)

2019 ◽  
Vol 10 (2) ◽  
pp. 117-124
Author(s):  
Nezahat Turfan ◽  
Sezgin Ayan ◽  
Esra Nurten Yer ◽  
Halil Barış Özel
Keyword(s):  
Phenolic Compounds ◽  
Soluble Protein ◽  
Soluble Sugars ◽  
Total Soluble Protein ◽  
Total Phenolic ◽  
Total Chlorophyll ◽  
H2o2 Concentration ◽  
Total Soluble Sugars ◽  
Oriental Beech ◽  
Different Ages

Background and Purpose: This study presents the analysis of photosynthetic pigments, proline, total soluble protein, total amino acids, glucose, sucrose, total soluble sugars, total amount of phenolic compounds and flavonoids, malondialdehyde (MDA) and hydrogen peroxide (H2O2) concentration in the leaf samples collected from oriental beech trees, which are naturally spread in Kastamonu Province, Turkey, with differing ages, enzyme activities of ascorbate peroxidase (APX), catalase (CAT) and superoxide dismutase (SOD). Material and Methods: The research was carried out on oriental beech trees (Fagus orientalis L.) of different ages located at 1300 m high elevation in Ahlat Village of Kastamonu Province, Turkey. Oriental beech trees of different ages (≥25, ≥50, ≥100, ≥200 and ≥600 years-old) constituted the material of this study. In leaf samples taken from trees of different ages, photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophyll and carotenoid), proline, total soluble protein, total amino acid, glucose, sucrose, total soluble sugars, the amount of total phenolic compounds and flavonoids, MDA, H2O2 concentration, enzyme activities of APX, CAT and SOD, as well as the relationship between the total content of C, N and H elements and the tree ages were studied. Results: As a result of the research conducted, significant differences were determined in terms of chlorophyll, total phenolic compound, flavonoid, glucose, amounts of sucrose, nitrogenous compounds, proline, total soluble protein, MDA, H 2O2 concentrations, and the activities of APX, CAT and SOD in the leaves of oriental beech trees with differing ages. The highest content of chlorophyll a was found to be in the youngest age group of ≥25 years. Total chlorophyll is low in young trees and high in middle-aged, old and very old trees. According to the results obtained, it was concluded that the MDA and H2O2 concentrations in the trees did not vary depending on the age of trees only, but also on the genotype, environmental conditions and metabolic activities. It was concluded that the fact that the total chlorophyll, phenolic compounds and sucrose content in oriental beech trees are high and that MDA content is low could have an influence on the long life of ≥600 years-old oriental beech trees. Conclusions: The activity of photosynthesis is related to leaf characteristics more than the age of trees.

scholarly journals Effect of Hydrolysis time and Papain Concentration on Some Properties of Apple Snail (Pilla ampullacea) Hydrolysate

2020 ◽  
Vol 1 (02) ◽  
pp. 1-5
Author(s):  
Andre Yusuf Trisna Putra ◽  
Dedin Finatsiyatull Rosida ◽  
Anugerah Dany Priyanto
Keyword(s):  
Soluble Protein ◽  
Enzyme Concentration ◽  
Total Soluble Protein ◽  
Apple Snail ◽  
Gut Content ◽  
Hydrolysis Time ◽  
Umami Taste ◽  
Nutritional Potential

content and total peptide content were investigated in the non-farmed freshwater apple snail (Pila ampullacea) to understand its nutritional potential as alternative umami taste enhancer. Apple Snail samples with removed gut content were collected from a local snail non-farm in Surabaya City. Papain enzyme was used. The two variables, hydrolysis time (3, 6, 9, 12, 15, 18 h) and enzyme concentration (1%, 5%, 10%), was used to produce the apple snail hydrolysate. The result showed that total soluble protein was about 2.1%-7.3%, whereas total peptide content was 5.05-15.73mg/ml. The highest total soluble protein was achieved at 12 hour. Longer hydrolysis time significantly effect on total peptide content of apple snail hydrolysate.

Methylene blue active substances in plaque of Bacillus subtilis subsp. subtilis and enrichment by supplemental calcium in culture media

2020 ◽  
Vol 71 (5) ◽  
pp. 550-556
Author(s):  
L.A. Krueger ◽  
M.A. Grzemski ◽  
M.C. Bilyeu ◽  
J.G. Horst ◽  
S.A. Ugrin ◽  
...  
Keyword(s):  
Methylene Blue ◽  
Bacillus Subtilis ◽  
Culture Media ◽  
Supplemental Calcium ◽  
Subtilis Subsp ◽  
Active Substances

scholarly journals Use of Biofuel Industry Wastes as Alternative Nutrient Sources for DHA-Yielding Schizochytrium limacinum Production

2020 ◽  
Vol 10 (12) ◽  
pp. 4398
Author(s):  
Sofoklis Bouras ◽  
Nikolaos Katsoulas ◽  
Dimitrios Antoniadis ◽  
Ioannis T. Karapanagiotidis
Keyword(s):  
Fatty Acid ◽  
Acid Content ◽  
Crude Glycerol ◽  
Sea Water ◽  
Culture Media ◽  
Fatty Acid Content ◽  
Nutrient Sources ◽  
Lipid Yield ◽  
Biofuel Industry ◽  
Schizochytrium Limacinum

The simultaneous use of crude glycerol and effluent from anaerobic digestate, both wastes derived from the biofuel industry, were tested in the frame of circular economy concept, as potential low-cost nutrient sources for the cultivation of rich in docosahexaenoic acid (DHA) oil microalgae strain Schizochytrium limacinum SR21. Initially, the optimal carbon and nitrogen concentration levels for high S. limacinum biomass and lipids production were determined, in a culture media containing conventional, high cost, organic nitrogen sources (yeast extract and peptone), micronutrients and crude glycerol at varying concentrations. Then, the effect of a culture media composed of crude glycerol (as carbon source) and effluent digestate at varying proportions on biomass productivity, lipid accumulation, proximate composition, carbon assimilation and fatty acid content were determined. It was shown that the biomass and total lipid content increased considerably with varying effluent concentrations reaching 49.2 g L−1 at 48% (v/v) of effluent concentration, while the lipid yield at the same effluent concentration reached 10.15 g L−1, compared to 17.0 g L−1 dry biomass and 10.2 g L−1 lipid yield when yeast extract and peptone medium with micronutrients was used. Compared to the control treatment, the above production was obtained with 48% less inorganic salts, which are needed for the preparation of the artificial sea water. It was shown that Schizochytrium limacinum SR21 was able to remediate 40% of the total organic carbon content of the biofuel wastes, while DHA productivity remained at low levels with saturated fatty acids comprising the main fraction of total fatty acid content. The results of the present study suggest that the simultaneous use of two waste streams from the biofuel industry can serve as potential nutrient sources for the growth of Schizochytrium limacinum SR21, replacing the high cost organic nutrients and up to one half the required artificial sea water salts, but upregulation of DHA productivity through optimization of the abiotic environment is necessary for industrial application, including aqua feed production.

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