Invertase Synthesis Research Articles

Multi-omics analysis of the regulatory effects of low-phosphorus stress on phosphorus transport in soybean roots.

The regulatory effects of uneven phosphorus supplies on phosphorus transport in soybean roots are still unclear. To further analyze the regulatory effects of low-phosphorus stress on phosphorus transport in soybean roots and the effects of uneven phosphorus application on the physiological mechanism of phosphorus transport in soybean roots, dual-root soybean plants were prepared via grafting, and a sand culture experiment was performed. From the unfolded cotyledon stage to the initial flowering stage, one side of each dual-root soybean system was irrigated with a low-phosphorus-concentration solution (phosphorus-application [P+] side), and the other side was irrigated with a phosphorus-free nutrient solution (phosphorus-free [P-] side); this setup allowed the study of the effects of different phosphorus supply levels on the expression of genes and proteins and the accumulation of metabolites in soybean roots on the P- side to clarify the method through which phosphorus transport is regulated in soybean roots and to provide a theoretical basis for improving the use rate of phosphorus fertilizer. The results revealed that the unilateral supply of low-concentration phosphorus promoted the uptake of phosphorus by soybean roots and the transport of phosphorus from the P+ side to the P- side. Compared with the normal concentration of phosphorus supply and the phosphorus-free supply, the low concentration phosphorus supply affected the regulation of the metabolic pathways involved in starch and sucrose metabolism, glycolysis, fructose, and mannose metabolism, etc., thereby affecting soybean root phosphorus transport. The low-phosphorus stress inhibited fructose synthesis and sucrose synthase synthesis in the soybean roots and the synthesis of hexokinase (HK) and fructose kinase, which catalyzes the conversion of fructose to fructose-6-phosphate. Low-phosphorus stress promoted the synthesis of sucrose invertase and the conversion of sucrose into maltose by the activity of starch synthase (StS) and stimulated the synthesis of UDPG pyrophosphorylase (UGP) and phosphoglucose isomerase (GP1), which is involved in the conversion of UDP-glucose to glucose-6-phosphate. The phosphorus transport pathway of soybean roots was then affected, which promoted phosphorus allocation to UTP and glucose-6-phosphate. Additionally, low-phosphorus stress hastened glycolysis in the soybean roots and inhibited the synthesis of malic acid, thereby promoting the transport of phosphorus in the roots. In addition, low-phosphorus stress inhibited the synthesis of fructose, mannose, and mannose-1-phosphate and the synthesis of other enzymes involved in phosphorus transport as well as invertase, thereby inhibiting the transport and synthesis of several organic phosphorus-containing compounds.

Invertase enzyme

The enzyme known as invertase catalysis the conversion of sucrose into glucose and fructose (EC 3.2.1.26). The best source of invertase is yeast. Internal and exterior in vertases are the two varieties found in S. cerevisiae. Invertase catalyses the hydrolysis of sucrose into fructose and glucose. Pharmaceuticals, analytical measuring probes, and yeast species detection ELISA kits all include this invertase. Immobilization of the invertase naturally is what is meant by this. Different levels of glycoprotein polymerization, glycosylation, and phosphorylation make bacteria invertase unique. A range of gram-positive and gram-negative organisms, including Zymomonas mobilis, and Bacillus strains, produce invertase. Comparing fungal growth to the time course of invertase synthesis by fungi, which ranged from 24 to 192 hours, was analysed. Twenty-four-hour intervals were used to measure the invertase activity. The biological characteristics that were looked at in this study included the amount of invertase-producing bacteria and soil invertase activity. Invertase is used to replace traditional acid hydrolysis.

SCREENING OF FILAMENTOUS FUNGI SYNTHESIZING INVERTASE

Results of screening invertase-synthesizing filamentous fungi deposited at Belarusian collection of non-pathogenic microorganisms and isolated from various sugar-containing substrates have been presented. Model organisms were selected to
 study the mechanisms of synthesis and properties of invertase.

Relationships of Light to Enzyme and Sucrose Transformations in Sugarcane Treated with Paraquat

Relationships of Light to Enzyme and Sucrose Transformations in Sugarcane Treated with Paraquat

Phenotypic regulation of the synthesis of carbohydrases by Kluyveromyces yeasts

The ability of Kluyveromyces marxianus Y-303 strain to synthesize highly active intracellular β-fructosidases (invertase and inulinase) was studied. The effect of separate sources of carbon, nitrogen, the initial pH of the nutrient medium, temperature and duration of cultivation process on the biosynthesis of enzymes were studied. The optimal composition of the nutrient medium and cultivation conditions that ensure the maximum production of the studied enzymes by K. marxianus yeast were selected. It was found out that the optimal medium for invertase synthesis contains 3% fructose and 0.6% NH4H2PO4, and for inulinase - 0.5% inulin and 0.12% yeast extract of inulinase. Optimal cultivation conditions for the accumulation of invertase - pH 6.0, temperature - 30 ° C, duration - 42 hours; and for inulinase, respectively: pH 5.0, 25 °C, 66 hours. The described cultivation conditions of K. marxianus led to 6-7 fold activity increase for invertaseand 8–9 fold- for inulinase.

Exogenous cytokinin application to Actinidia chinensis var. deliciosa \u2018Hayward\u2019 fruit promotes fruit expansion through water uptake

Exogenous application of a cytokinin-like compound forchlorfenuron (CPPU) can promote fruit growth, although often at the expense of dry matter (DM), an important indicator of fruit quality. Actinidia chinensis var. deliciosa ‘Hayward’ fruit are very responsive to CPPU treatments, but the mechanism underlying the significant fruit weight increase and associated decrease in DM is unclear. In this study, we hypothesised that CPPU-enhanced growth increases fruit carbohydrate demand, but limited carbohydrate supply resulted in decreased fruit DM. During fruit development, CPPU effects on physical parameters, metabolites, osmotic pressure and transcriptional changes were assessed under conditions of both standard and a high carbohydrate supply. We showed that CPPU increased fruit fresh weight but the dramatic DM decrease was not carbohydrate limited. Enhanced glucose and fructose concentrations contributed to an increase in soluble carbohydrate osmotic pressure, which was correlated with increased water accumulation in CPPU-treated fruit and up-regulation of water channel aquaporin gene PIP2.4 at 49 days after anthesis. Transcipt analysis suggested that the molecular mechanism contributing to increased glucose and fructose concentrations was altered by carbohydrate supply. At standard carbohydrate supply, the early glucose increase in CPPU fruit was associated with reduced starch synthesis and increased starch degradation. When carbohydrate supply was high, the early glucose increase in CPPU fruit was associated with a general decrease in starch synthesis but up-regulation of vacuolar invertase and fructokinase genes. We conclude that CPPU affected fruit expansion by increasing the osmotically-driven water uptake and its effect was not carbohydrate supply-limited.

Improvement of Invertase Synthesis by the Mutant Saccharomyces cerevisiae through UV Mutagenesis

In fermentation technology, strain improvement of baker's yeast has traditionally relied on random mutagenesis followed by screening for mutant exhibiting enhanced properties of interest. Such mutant organisms are useful in several industries. Saccharomyces cerevisiae can use sucrose as the sole source of both carbon and energy; hydrolysis of this sugar is catalyzed by the enzyme invertase. The main objective of this work is to overcome the glucose repression of invertase by invertase constitutive mutants through UV mutation. This may occur in any glucose repressible genes as a single or double mutation in repressor gene (s) which might cause constitutive synthesis of invertase. These mutated screened strains were optimized with various glucose concentration and different incubation hours for higher invertase production. The maximum synthesis of invertase was 1.066 units/ml in hour from mutant Saccharomyces cerevisiae type-2 strain.

Effects of nitrogen and carbon sources on the production of inulinase from strain Bacillus sp. SG113

Abstract The effects of the carbon and nitrogen substrates on the growth of Bacillus sp. SG113 strain were studied. The use of organic nitrogen sources (peptone, beef extract, yeast extract, casein) leads to rapid cellular growth and the best results for the Bacillus strain were obtained with casein hydrolysate. From the inorganic nitrogen sources studied, the (NH4) 2SO4 proved to be the best nitrogen source. Casein hydrolysate and (NH4) 2SO4 stimulated the invertase synthesis. In the presence of Jerusalem artichoke, onion and garlic extracts as carbon sources the strain synthesized from 6 to 10 times more inulinase.

Invertase-nanogold clusters decorated plant membranes for fluorescence-based sucrose sensor.

In the present study, invertase-mediated nanogold clusters were synthesized on onion membranes, and their application for sucrose biosensor fabrication was investigated. Transmission electron microscopy revealed free nanoparticles of various sizes (diameter ~5 to 50 nm) along with clusters of nanogold (~95 to 200 nm) on the surface of inner epidermal membranes of onions (Allium cepa L.). Most of the polydispersed nanoparticles were spherical, although some were square shaped, triangular, hexagonal or rod-shaped. Ultraviolet–visible spectrophotometric observations showed the characteristic peak for nanoparticles decorated invertase-onion membrane at approximately 301 nm. When excited at 320 nm in the presence of sucrose, the membranes exhibited a photoemission peak at 348 nm. The fluorescence lifetime of this nanogold modified onion membrane was 6.20 ns, compared to 2.47 ns for invertase-onion membrane without nanogold. Therefore, a sucrose detection scheme comprised of an invertase/nanogold decorated onion membrane was successfully developed. This fluorescent nanogold-embedded onion membrane drop-test sensor exhibited wide acidic to neutral working pH range (4.0-7.0) with a response time 30 seconds (<1 min). The fabricated quenching-based probe had a low detection limit (2x10−9 M) with a linear dynamic range of 2.25x10−9 to 4.25x10−8 M for sensing sucrose. A microplate designed with an enzyme-nanomaterial-based sensor platform exhibited a high compliance, with acceptable percentage error for the detection of sucrose in green tea samples in comparison to a traditional method. With some further, modifications, this fabricated enzyme-nanogold onion membrane sensor probe could be used to estimate glucose concentrations for a variety of analytical samples.Graphical abstractSynthesis and characterization of invertase assisted nanogold clusters on onion membranes and their application for fluorescence-based sucrose sensor.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0089-1) contains supplementary material, which is available to authorized users.

Kinetic and Energetic Parameters of Carob Wastes Fermentation by Saccharomyces cerevisiae: Crabtree Effect, Ethanol Toxicity, and Invertase Repression.

Carob waste is a useful raw material for the second-generation ethanol because 50% of its dry weight is sucrose, glucose, and fructose. To optimize the process, we have studied the influence of the initial concentration of sugars on the fermentation performance of Saccharomyces cerevisiae. With initial sugar concentrations (S0) of 20 g/l, the yeasts were derepressed and the ethanol produced during the exponential phase was consumed in a diauxic phase. The rate of ethanol consumption decreased with increasing S0 and disappeared at 250 g/l when the Crabtree effect was complete and almost all the sugar consumed was transformed into ethanol with a yield factor of 0.42 g/g. Sucrose hydrolysis was delayed at high S0 because of glucose repression of invertase synthesis, which was triggered at concentrations above 40 g/l. At S0 higher than 250 g/l, even when glucose had been exhausted, sucrose was hydrolyzed very slowly, probably due to an inhibition at this low water activity. Although with lower metabolic rates and longer times of fermentation, 250 g/l is considered the optimal initial concentration because it avoids the diauxic consumption of ethanol and maintains enough invertase activity to consume all the sucrose, and also avoids the inhibitions due to lower water activities at higher S0.

Improvement of l-lactic acid production from Jerusalem artichoke tubers by mixed culture of Aspergillus niger and Lactobacillus sp.

Aspergillus niger SL-09 and Lactobacillus sp. G-02 were used as a mixed culture in a 7-l fermentor to directly form l-lactic acid from Jerusalem artichoke tubers. The synthesis of inulinase and invertase from A. niger SL-09 was enhanced significantly by the inoculation of Lactobacillus sp. G-02 at 12 h of culture, which reached 275.6 and 571.8 U/ml in 60 h, over 5-folds higher than that of the culture using single strain. In the following simultaneous saccharification and fermentation procedure, the highest l-lactic acid concentration of 120.5 g/l was obtained in 36 h of the fed-batch fermentation with high conversion efficiency of 94.5%.

Regulation of invertase synthesis in Aspergillus niger

The regulation of the synthesis of invertase in Aspergillus niger with different carbohydrates was studied. In order to investigate the mechanism of induction, we worked with conidia protoplasts and permeabilied mycelium and used actinomycin D, cycloheximide and cAMP. The synthesis of invertase in A. niger was induced by β-fructofuranoside substrates such as raffinose, sucrose and turanose. Inulin also produced synthesis, suggesting that the β link and the fructose located at the end of the molecule are involved in the induction mechanism. Glucose and fructose, which are products of the action of invertase, are repressors of the synthesis and act at the level of the translation of the genetic message. The induction of invertase is different from that of yeasts and other filamentous fungi. A mechanism of induction of the synthesis of invertase in A. niger with the participation of cAMP is proposed.

Effect of osmotic stress on the derepression of invertase synthesis in nonconventional yeasts

The aim of this study was to analyse the effect of osmotic stress on the biosynthesis of invertase enzyme in nonconventional yeasts. Invertase activities of the nonconventional yeast species belonging to Kluyveromyces, Schwanniomyces and Pichia genus were measured either in the presence or in the absence of various amounts of NaCl. The effect of hyperosmotic stress on the glucose consumption of Saccharomyces cerevisiae and Pichia anomala were also compared. Like S. cerevisiae, derepression of invertase synthesis in Kluyveromyces lactis, Schwanniomyces occidentalis and Pichia jadinii is inhibited by hyperosmotic stress. However, derepression of invertase synthesis in P. anomala is not affected by hyperosmotic stress. In addition, low levels of osmotic stress activated invertase synthesis three- to fourfold in P. anomala and K. lactis. This study shows that low levels of osmotic stress induces the invertase synthesis at very high levels in P. anomala and K. lactis. Glucose consumption was not influenced at significant levels by the hyperosmotic stress in P. anomala. This study shows the activation of invertase synthesis by low levels of osmotic stress in P. anomala and K. lactis.

Effect of cultivation conditions on invertase production by hyperproducing Saccharomyces cerevisiae isolates

Invertase (β-D-fructofuranoside fructohydrolase, EC 3.2.1.26) finds major uses in confectionery and in the production of invert syrup. In the present study, we report on invertase production by wild cultures of Saccharomyces cerevisiae. The yeast strains were isolated from dates available in a local market. Five hyperproducing yeast strains (>100- fold higher invertase activity) were kinetically analysed for invertase production. Saccharomyces cerevisiae strain GCA-II was found to be a better invertase-yielding strain than all the other isolates. The values of Qp and Yp/s for GCA-II were economical as compared to other Saccharomyces cultures. The effect of sucrose concentration, rate of invertase synthesis, initial pH of fermentation medium and different organic nitrogen sources on the production of invertase under submerged culture conditions was investigated. Optimum concentrations of sucrose, urea and pH were 3, 0.2 (w/v), and 6 respectively. The increase in the enzyme yield obtained after optimization of the cultural conditions was 47.7%.

Non-Histone Proteins Nhp6A and Nhp6B Are Required for the Regulated Expression of SUC2 Gene of Saccharomyces cerevisiae

Transcription of the SUC2 gene that encodes invertase enzyme is controlled by glucose repression and derepression mechanisms in Saccharomyces cerevisiae. Several regulatory factors such as Mig1p complex, Gcr1p, Hxk2p, nucleosomes, and the Snf1p kinase complex have been identified as the regulators of SUC2 transcription. The results presented in this study indicate that the non-histone proteins Nhp6A and Nhp6B were also required for the regulated expression of SUC2 gene. Expression of the SUC2 gene reduced to one-fiftieth–one-tenth in the Δnhp6A Δnhp6B double mutant strain depending on the growth conditions. Moreover, SUC2 expression and invertase synthesis became constitutive after long-term derepression, and decreased to a low level in Δnhp6A Δnhp6B double deletion mutant. A time course analysis of the invertase synthesis revealed that both the repression and derepression rates were very slow in the Δnhp6A Δnhp6B double mutant yeast. These results indicate that the architectural transcription factors Nhp6A and Nhp6B play a very critical role in the regulation of SUC2 gene expression.

Cloning and characterization of glucokinase from a methylotrophic yeast Hansenulapolymorpha: different effects on glucose repression in H. polymorpha and Saccharomycescerevisiae

Glucokinase gene (HPGLK1) was cloned from a methylotrophic yeast Hansenulapolymorpha by complementation of glucose-phosphorylation deficiency in a H. polymorpha double kinase-negative mutant A31-10 by a genomic library. An open reading frame of 1416 nt encoding a 471-amino-acid protein with calculated molecular weight 51.6 kDa was characterized in the genomic insert of the plasmid pH3. The protein sequence deduced from HPGLK1 exhibited 55 and 46% identity with glucokinases from Saccharomycescerevisiae and Aspergillusniger, respectively. The enzyme phosphorylated glucose, mannose and 2-deoxyglucose, but not fructose. Transformation of HPGLK1 into A31-10 restored glucose repression of alcohol oxidase and catalase in the mutant. Transformation of HPGLK1 into S. cerevisiae triple kinase-negative mutant DFY632 showed that H. polymorpha glucokinase cannot transmit the glucose repression signal in S. cerevsiae: synthesis of invertase and maltase in respective transformants was insensitive to glucose repression similarly to S. cerevisiae DFY568 possessing only glucokinase.

Characterization of a mutant from Lactobacillus amylovorus JCM 1126(T) with improved utilization of sucrose.

A strain YF43, which can grow on sucrose as rapidly as glucose, was isolated by mutation from Lactobacillus amylovorus JCM 1126, the type strain defective in sucrose utilization. Exogenous sucrose stimulated the production of invertase by strains YF43 and JCM 1126 simultaneously. In a medium containing fructooligosaccharide as the sole carbon source, the cells of strain YF43 showed high invertase activity in spite of poor growth. The two invertases produced in the cells grown on sucrose and fructooligosaccharide were an identical beta-fructofuranosidase, as judged from properties of partially purified enzymes. These observations indicated that strain YF43 is a mutant improved for permeation of sucrose and not derepressed for the synthesis of invertase.

Gibberellin‐induced elongation and osmoregulation in internodes of floating rice

Internodal elongation in floating rice (Oryza sativa L. cv. Habiganj Aman II) is known to be enhanced by treatment with ethylene or gibberellic acid (GA3) at high relative humidity (RH). However, ethylene‐induced internodal elongation is inhibited at low RH. while GA3‐induced internodal elongation is hardly affected by humidity. We examined the possible involvement of osmoregulation in the stimulation by GA3 of the elongation of internodes at low RH. Submergence and treatment with ethylene or GA33 at 100% RH increased the osmotic potential in internodes of excised stem segments, while GA3 at 20% RH maintained the osmotic potential at a low level. In internodes of stem segments that had been treated with GA3 at 20% RH, the activity of invertase and the level of soluble sugars were almost 2‐ and 1.5‐fold higher, respectively, than those in internodes that had been treated with GA3 at 100% RH. These results indicate that one of the possible mechanisms by which GA3 promotes elongation of internodes at low RH involves the osmoregulation that is achieved by promotion of the synthesis of invertase.

Gibberellin-induced elongation and osmoregulation in internodes of floating rice

Internodal elongation in floating rice (Oryza sativa L. cv. Habiganj Aman II) is known to be enhanced by treatment with ethylene or gibberellic acid (GA 3 ) at high relative humidity (RH). However, ethylene-induced internodal elongation is inhibited at low RH, while GA 3 -induced internodal elongation is hardly affected by humidity. We examined the possible involvement of osmoregulation in the stimulation by GA 3 of the elongation of internodes at low RH. Submergence and treatment with ethylene or GA 3 at 100% RH increased the osmotic potential in internodes of excised stem segments, while GA 3 at 20% RH maintained the osmotic potential at a low level. In internodes of stem segments that had been treated with GA 3 at 20% RH, the activity of invertase and the level of soluble sugars were almost 2- and 1.5-fold higher, respectively, than those in internodes that had been treated with GA 3 at 100% RH. These results indicate that one of the possible mechanisms by which GA 3 promotes elongation of internodes at low RH involves the osmoregulation that is achieved by promotion of the synthesis of invertase.

Purification and characterization of an invertase from Candida utilis: comparison with natural and recombinant yeast invertases

A periplasmic invertase from the yeast Candida utilis was purified to homogeneity from cells fully derepressed for invertase synthesis. The enzyme was purified by successive Sephacryl S-300, and affinity chromatography and shown to be a dimeric glycoprotein composed of two identical monomer subunits with an apparent molecular mass of 150 kDa. After EndoH treatment, the deglycosylated protein showed an apparent molecular weight of 60 kDa. The apparent K m values for sucrose and raffinose were 11 and 150 mM, respectively, similar to those reported in Saccharomyces cerevisiae. The range of optimum temperature was 60–75°C. The optimum pH was 5.5 and the enzyme was stable over pH range 3–6.