共查询到20条相似文献,搜索用时 31 毫秒
1.
Yasutake N Totani K Harada Y Haraguchi S Murata T Usui T 《Bioscience, biotechnology, and biochemistry》2003,67(7):1530-1536
A condensation reaction between N-acetyllactosamine and glycerol was directly catalyzed by using a commercially available cellulase preparation from Trichoderma reesei. 1-O-beta-N-Acetyllactosaminyl-(R, S)-glycerols (1) were readily synthesized in a 5% yield based on the N-acetyllactosamine added and conveniently isolated by two-step column chromatographies. The use of a partially purified enzyme increased 2.3-fold the yield of 1, compared to that of the crude enzyme containing beta-D-galactosidase activity. When various alkanols (n:2-4) were used in the condensation reaction, the corresponding alkyl beta-N-acetyllactosaminides were obtained in yields of 0.3-1.1% of the desired compounds. 相似文献
2.
《Bioscience, biotechnology, and biochemistry》2013,77(7):1530-1536
A condensation reaction between N-acetyllactosamine and glycerol was directly catalyzed by using a commercially available cellulase preparation from Trichoderma reesei. 1-O-β-N-Acetyllactosaminyl-(R, S)-glycerols (1) were readily synthesized in a 5% yield based on the N-acetyllactosamine added and conveniently isolated by two-step column chromatographies. The use of a partially purified enzyme increased 2.3-fold the yield of 1, compared to that of the crude enzyme containing β-D-galactosidase activity. When various alkanols (n:2-4) were used in the condensation reaction, the corresponding alkyl β-N-acetyllactosaminides were obtained in yields of 0.3-1.1% of the desired compounds. 相似文献
3.
Effect of lactose and glycerol on the motility, normal apical ridge, chromatin condensation and chromatin stability of frozen boar spermatozoa 总被引:1,自引:0,他引:1
Corcuera BD Marigorta P Sagüés A Saiz-Cidoncha F Pérez-Gutiérrez JF 《Theriogenology》2007,67(6):1150-1157
The effect of lactose and glycerol concentration, as well as the equilibration time with glycerol was studied on motility, normal apical ridge (NAR), and chromatin state of boar spermatozoa after the freezing and thawing process. In the first experiment, samples were frozen in first and second extenders containing different concentrations of lactose (11, 12 and 14%). In the second experiment samples were frozen using second extenders with different concentrations of glycerol (4, 6, 8 and 10%) and were incubated at 5 degrees C for 0 and 30 min. Motility, motility after caffeine treatment, NAR, chromatin condensation and stability (susceptibility to de-condense after heparin treatment) were evaluated. The results indicated that freezing spermatozoa in extenders with increasing concentrations of lactose adversely affected motility but provided a protective effect on acrosomes. Increased lactose concentration induced higher chromatin condensation but maintained the same stability. Increasing the glycerol concentration in the second extender from 4-6 to 8% led to higher motility and NAR as well as lower chromatin condensation and stability. When 30 min equilibration time was allowed after dilution with the same extenders, spermatozoa showed higher NAR and lower chromatin condensation and stability. The longer equilibration time was detrimental for motility when freezing in the 8% glycerol extender but favourable when using the 4% glycerol extender. Compared to the 8% glycerol, spermatozoa frozen in the 10% glycerol extender showed similar motility and increased chromatin condensation and stability, as well as low values of NAR that did not improve by longer incubation time. 相似文献
4.
Purification and biochemical properties of a galactooligosaccharide producing β-galactosidase from Bullera singularis 总被引:2,自引:0,他引:2
A beta-galactosidase, catalyzing lactose hydrolysis and galactooligosaccharide (GalOS) synthesis from lactose, was extracted from the yeast, Bullera singularis KCTC 7534. The crude enzyme had a high transgalactosylation activity resulting in the oligosaccharide conversion of over 34% using pure lactose and cheese whey permeate as substrates. The enzyme was purified by two chromatographic steps giving 96-fold purification with a yield of 16%. The molecular weight of the purified enzyme (specific activity of 56 U mg(-1)) was approx. 53 000 Da. The hydrolytic activity was the highest at pH 5 and 50 degrees C, and was stable to 45 degrees C for 2 h. Enzyme activity was inhibited by 10 mM Ag3+ and 10 mM SDS. The Km for lactose hydrolysis was 0.58 M and the maximum reaction velocity (V(max)) was 4 mM min(-1). GalOS, including tri- and tetra-saccharides were produced with a conversion yield of 50%, corresponding to 90 g GalOS l(-1) from 180 g lactose l(-1) by the purified enzyme. 相似文献
5.
The effect of enzyme to substrate ratio, initial lactose concentration and temperature has been studied for the kinetically controlled reaction of lactose transgalactosylation with Aspergillus oryzae β-galactosidase, to produce prebiotic galacto-oligosaccharides (GOS). Enzyme to substrate ratio had no significant effect on maximum yield and specific productivity. Galacto-oligosaccharide syntheses at very high lactose concentrations (40, 50 and 60%, w/w, lactose monohydrate) were evaluated at different temperatures (40, 47.5 and 55°C). Within these ranges, lactose could be found as a supersaturated solution or a heterogeneous system with precipitated lactose, resulting in significant effect on GOS synthesis. An increase in initial lactose concentration produced a slight increase in maximum yield as long as lactose remained dissolved. Increase in temperature produced a slight decrease in maximum yield and an increase in specific productivity when supersaturation of lactose occurred during reaction. Highest yield of 29 g GOS/100 g lactose added was obtained at a lactose monohydrate initial concentration of 50% (w/w) and 47.5°C. Highest specific productivity of 0.38 g GOSh(-1) mg enzyme(-1) was obtained at lactose monohydrate initial concentration of 40% (w/w) and 55°C, where a maximum yield of 27 g GOS/100 g lactose added was reached. This reflects the complex interplay between temperature and initial lactose concentration on the reaction of synthesis. When lactose precipitation occurred, values of yields and specific productivities lower than 22 g GOS/100 g lactose added and 0.03 gGOSh(-1) mg enzyme(-1) were obtained, respectively. 相似文献
6.
Sapna Das-Bradoo Ingemar Svensson Jorge Santos Fatima Plieva Bo Mattiasson Rajni Hatti-Kaul 《Journal of biotechnology》2004,110(3):273-286
Whole cells of alkaliphilic Bacillus pseudofirmus AR-199, induced for β-galactosidase activity, were used for the synthesis of 1-hexyl-β-
-galactoside and 1-octyl-β-
-galactoside, respectively, by transglycosylation reaction between lactose and the corresponding alcohol acceptor. The product yield was strongly influenced by the initial water content in the reaction mixture. Water content of 10% (v/v) was optimal providing 3.6–36 mM hexyl galactoside from 10 to 150 mM lactose, and no secondary product hydrolysis. Product yield could be enhanced by supplementing the reaction mixture with more cells or partly replacing the product with fresh substrate, but was decreased with time to the initial equilibrium level. Cell permeabilisation or disruption resulted in increased reaction rate and higher product yield but was followed by product hydrolysis. Octyl galactoside synthesis using whole cells was optimal at water content of 2% (v/v) with a yield of 26%. The cells were immobilised in cryogels of polyvinyl alcohol for use in continuous process, where hexyl galactoside was produced with a constant yield of 50% from 50 mM lactose for at least a week. 相似文献
7.
The production of a neutral lipase from a Bacillus sp. was improved tremendously (193-fold) following media optimization involving both the "one-at-a-time" and the statistical designing approaches. The present lipase was poorly induced by oils, instead its production was induced in the presence of sugars and sugar alcohols, mainly galactose, lactose, glycerol, and mannitol. A high inoculum density of 15% v/v (A550 = 0.8) led to maximum lipase production. Interestingly, the enzyme induction was growth independent, a property very different from most of the lipases investigated to date. The optimal composition of the growth medium to achieve maximum lipase production was determined to be as follows: NH4Cl, 35 g x L(-1); glycerol, 10 mL x L(-1); K2HPO4, 3 g x L(-1); KH2PO4, 1 g x L(-1); MgSO4.7H2O, 0.1 g x L(-1); glucose, 2 g x L(-1); MgCl2, 0.6 mmol x L(-1), with 15% inoculum density and an incubation period of 24 h. About 62 U x mL(-1) of enzyme production was achieved in the optimized medium. 相似文献
8.
Niiyama K Yoshizumi T Takahashi H Naya A Ohtake N Fukami T Mase T Hayama T Ishikawa K 《Bioorganic & medicinal chemistry》2002,10(11):3437-3444
A convenient method for the synthesis of the title intermediate 4 was described. The key steps of this synthesis involved: (1) regioselective addition reaction of arylzinc reagent to quinolic anhydride in 42% isolated yield, (2) conversion of a ketoacid to an enone, which was achieved in 65% yield by intramolecular Knoevenagel reaction of beta-ketoester generated by condensation of an acid imidazolide with an ester enolate, followed by dehydration assisted with silica gel, and (3) stereoselective reduction of an allyl alcohol in 75% yield with zinc under acidic conditions. This synthesis enabled us to provide hundreds of grams of without chromatographic purification. 相似文献
9.
Koto S Hirooka M Tashiro T Sakashita M Hatachi M Kono T Shimizu M Yoshida N Kurasawa S Sakuma N Sawazaki S Takeuchi A Shoya N Nakamura E 《Carbohydrate research》2004,339(14):2415-2424
Alkyl, cycloalkyl, allyl, 4-pentenyl, and benzyl alpha-glycosides of maltose, cellobiose, and lactose were prepared (17-77% yield; alpha/beta=70/30-96/4) via a direct reaction of the free disaccharides with a binary AcBr-AcOH mixture, followed by glycosidation with alcohol using FeCl3 in MeNO2 or CH2Cl2, Zemplén deacetylation, and resolution of the anomeric mixture of glycosides by chromatography. Using MeCN as solvent for the glycosidation step, the corresponding beta-biosides were also prepared (16-61% yield; alpha/beta=25/75-5/95). 相似文献
10.
The production of extracellular 1,3-, 1,6-beta-glucanases and chitinase was studied during submerged cultivation of a Trichoderma viride strain 3/78 on various carbon sources: glycerol, glucose, lactose, sucrose, laminaran, starch, pustulan, chitin, and Agaricus bisporus fruit bodies. The synthesis of these enzymes and cellulase was studied also under the conditions of depression at low concentrations (10(-2) and 10(-3)M) of the first five aforementioned carbon sources as well as cellobiose, gentiobiose, N-acetyl-beta-D-glucosamine and 0.1% chitooligosaccharides and A. bisporus cell walls. The experiments were conducted with the washed mycelium of this strain grown for 2 days in a medium with glycerol as a carbon source. The results indicated that 1,3- and 1,6-beta-glucanases of the strain were of the constitutive nature and were repressed by such carbon sources as glycerol and glucose. Chitinase and cellulase were shown to be inducible enzymes. Chitinase was induced by N-acetyl-beta-D-glucosamine, chitooligosaccharides and A. bisporus cell walls as well as by lactose when the fungus was grown on this carbon source. Cellulase biosynthesis was induced by lactose, cellobiose and gentiobiose. 相似文献
11.
Harland G. Wood Georges J. Peeters Roger Verbeke Monique Lauryssens Birgit Jacobson 《The Biochemical journal》1965,96(3):607-615
1. The distributions of (14)C have been compared in the glucose and galactose moieties of lactose obtained from cows' udders perfused with blood containing [1-(14)C]-, [2-(14)C]- and [6-(14)C]-glucose. The (14)C of the glucose moiety was found in the same position as that of the administered glucose, but in the galactose moiety the (14)C from [2-(14)C]glucose was extensively randomized into positions 1 and 3. It is concluded that the glucose moiety arose from free glucose and the galactose moiety from hexose phosphate intermediates and that the latter reflected the randomization occurring through reactions of the pentose cycle. 2. The proportion of the glucose metabolized via the pentose cycle for those cells making lactose was estimated from the distribution of (14)C in the galactose moiety and found to be about 23% in one experiment and 30% in another experiment. 3. The yield and distribution of (14)C were determined in the glycerol of fat from the tissue in experiments with [2-(14)C]- and [6-(14)C]-glucose. There was a greater randomization of (14)C in the glycerol than in C-1, C-2 and C-3 of the galactose moiety of lactose. The ratio of the yield of (14)C in the glycerol from [2-(14)C]glucose to that of [6-(14)C]glucose was very low and from this ratio it was calculated that less than 10% of the glucose was metabolized by the Embden-Meyerhof pathway and approx. 60-70% was converted into lactose. 4. [6-(14)C]Glucose and [6-(3)H]glucose were used to determine whether the (3)H at the C-6 position remained stable during its conversion into glyceride of fat from the tissue. Twenty-seven per cent of the (3)H was labilized during this conversion. Therefore it was not possible to use [2-(14)C]glucose and [6-(3)H]glucose in a single experiment to measure the relative conversion of the C-2 and C-6 positions of glucose to glycerol. 相似文献
12.
Improvement of covalent immobilization procedure of β‐galactosidase from Kluyveromyces lactis for galactooligosaccharides production: Modeling and kinetic study
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Flor González‐Cataño Luz Tovar‐Castro Eduardo Castaño‐Tostado Carlos Regalado‐Gonzalez Blanca García‐Almendarez Anaberta Cardador‐Martínez Silvia Amaya‐Llano 《Biotechnology progress》2017,33(6):1568-1578
Galactooligosaccharides (GOS) are prebiotics produced from lactose through an enzymatic reaction. Employing an immobilized enzyme may result in cost reductions; however, the changes in its kinetics due to immobilization has not been studied. This study experimentally determined the optimal reaction conditions for the production of GOS from lactose by β‐galactosidase (EC 3.2.1.23) from Kluyveromyces lactis covalently immobilized to a polysiloxane‐polyvinyl alcohol (POS‐PVA) polymer activated with glutaraldehyde (GA), and to study the transgalactosylation kinetics. Yield immobilization was 99 ± 1.1% with 78.5 ± 2.4% enzyme activity recovery. An experimental design 24 with 1 center point and 2 replicates was used. Factors were lactose [L], enzyme concentration [E], pH and temperature (T). Response variables were glucose and galactose as monosaccharides [G1], residual lactose [Lac]r and GOS as disaccharides [G2] and trisaccharides [G3]. Best conditions were pH 7.1, 40 °C, 270 gL?1 initial lactose concentration and 6 U mL?1 enzyme concentration, obtaining 25.46 ± 0.01 gL?1 yield of trisaccharides. Although below the HPLC‐IR detection limit, tetrasaccharides were also identified after 115 min of reaction. The immobilization protocol was then optimized by diminishing total reactant volumes : support ratio, resulting in improved enzyme activity synthesizing 43.53 ± 0.02 gL?1 of trisaccharides and 13.79 ± 0.21 gL?1 of tetrasaccharides, and after four cycles remaining relative activity was 94%. A reaction mechanism was proposed through which a mathematical model was developed and rate constants were estimated, considering a pseudo steady‐state hypothesis for two concomitant reactions, and from this simplified analysis, the reaction yield could eventually be improved. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1568–1578, 2017 相似文献
13.
An alpha-L-fucosidase from porcine liver produced alpha-L-Fuc-(1-->2)-beta-D-Gal-(1-->4)-D-GlcNAc (2'-O-alpha-L-fucosyl-N-acetyllactosamine, 1) together with its isomers alpha-L-Fuc-(1-->3)-beta-D-Gal-(1-->4)-D-GlcNAc (2) and alpha-L-Fuc-(1-->6)-beta-D-Gal-(1-->4)-D-GlcNAc (3) through a transglycosylation reaction from p-nitrophenyl alpha-L-fucopyranoside and beta-D-Gal-(1-->4)-D-GlcNAc. The enzyme formed the trisaccharides 1-3 in 13% overall yield based on the donor, and in the ratio of 40:37:23. In contrast, transglycosylation by Alcaligenes sp. alpha-L-fucosidase led to the regioselective synthesis of trisaccharides containing a (1-->3)-linked alpha-L-fucosyl residue. When beta-D-Gal-(1-->4)-D-GlcNAc and lactose were acceptors, the enzyme formed regioselectively compound 2 and alpha-L-Fuc-(1-->3)-beta-D-Gal-(1-->4)-D-Glc (3'-O-alpha-L-fucosyllactose, 4), respectively, in 54 and 34% yields, based on the donor. 相似文献
14.
Optimization of hexyl-g-glycoside synthesis from lactose in hexanol at low water activity and high temperature was investigated using g-glycosidases from hyperthermophilic organisms: Sulfolobus solfataricus (LacS) and Pyrococcus furiosus (CelB). The method for water activity adjustment by equilibration with saturated salt solutions was adapted for use at high temperature. The influence of enzyme immobilization (on XAD-4, XAD-16, or Celite), addition of surfactants (AOT or SDS), substrate concentration, water activity, and temperature (60-90°C) on enzymatic activity and hexyl-g-glycoside yield were examined. Compared to other g-glycosidases in lactose conversion into alkyl glycoside, these enzymes showed high activity in a hexanol one-phase system and synthesized high yields of both hexyl-g-galactoside and hexyl-g-glucoside. Using 32 λg/l lactose (93 λmM), LacS synthesized yields of 41% galactoside (38.1 λmM) and 29% glucoside (27.0 λmM), and CelB synthesized yields of 63% galactoside (58.6 λmM) and 28% glucoside (26.1 λmM). With the addition of SDS to the reaction it was possible to increase the initial reaction rate of LacS and hexyl-g-galactoside yield (from 41 to 51%). The activity of the lyophilized enzyme was more influenced by the water content in the reaction than the enzyme on solid support. In addition, it was concluded that for the lyophilized enzyme preparation the enzymatic activity was much more influenced by the temperature when the water activity was increased. A variety of different glycosides were prepared using different alcohols as acceptors. 相似文献
15.
Das-Bradoo S Svensson I Santos J Plieva F Mattiasson B Hatti-Kaul R 《Journal of biotechnology》2004,110(3):273-285
Whole cells of alkaliphilic Bacillus pseudofirmus AR-199, induced for beta-galactosidase activity, were used for the synthesis of 1-hexyl-beta-d-galactoside and 1-octyl-beta-d-galactoside, respectively, by transglycosylation reaction between lactose and the corresponding alcohol acceptor. The product yield was strongly influenced by the initial water content in the reaction mixture. Water content of 10% (v/v) was optimal providing 3.6-36 mM hexyl galactoside from 10 to 150 mM lactose, and no secondary product hydrolysis. Product yield could be enhanced by supplementing the reaction mixture with more cells or partly replacing the product with fresh substrate, but was decreased with time to the initial equilibrium level. Cell permeabilisation or disruption resulted in increased reaction rate and higher product yield but was followed by product hydrolysis. Octyl galactoside synthesis using whole cells was optimal at water content of 2% (v/v) with a yield of 26%. The cells were immobilised in cryogels of polyvinyl alcohol for use in continuous process, where hexyl galactoside was produced with a constant yield of 50% from 50mM lactose for at least a week. 相似文献
16.
A novel lactose induction strategy for the production of rhLK68, the kringle fragments of human apolipoprotein(a) (apo(a)) as a novel anti-angiogenic protein, was investigated. A scale-up of the production was accompanied by a decrease in expression level, and severe aggregation occurred during the solubilization of rhLK68 from the inclusion body during a conventional single introduction of lactose. To overcome this problem, a continuous induction strategy was applied where lactose was mixed with glycerol and fed continuously in a dissolved oxygen (DO)-stat manner. With the sub-optimal feed medium consisted of 1:50 of lactose/glycerol (w/w), the expression level reached 16% of the total cellular protein, which was 1.6-fold higher than that obtained from the conventional lactose induction. Moreover, the solubilization yield of rhLK68 from the inclusion body increased from 30 +/- 5 to 85 +/- 3% compared to the conventional single introduction of lactose. This result suggests that the continuous lactose induction strategy beneficially influenced the expression level of rhLK68 and the quality of its inclusion body. 相似文献
17.
固定化嗜热脂肪芽孢杆菌合成低聚半乳糖 总被引:7,自引:2,他引:7
利用海藻酸钙、明胶和壳聚糖为固定化载体包埋嗜热脂肪芽孢杆菌细胞合成低聚半乳糖 (GOS)。通过比较三种方法的酶活力回收、最适反应条件、GOS的得率和和载体机械强度 ,选择明胶作为固定化细胞的载体。反应体系的温度、pH、乳糖浓度、乳糖的转化率和载体的传质阻力对GOS合成有明显影响。在CSTR反应器中水解 60 %乳糖 ,GOS最大得率为31 2 % ,经过 96h( 8批反应 ) ,产物得率为原来的 88%。在空速 0 0 9h- 1条件下 ,利用填充床反应器连续水解乳糖 ,GOS的得率和反应器生产能力分别为 31 5%和 1 7 4g (L·h) ,连续反应1 40h,GOS得率下降 2 0 %。产物经过活性炭柱层柱分离纯化 ,通过13C NMR鉴定四糖的化学结构为 β D Gal ( 1→ 3) D Gal ( 1→ 6) D G ( 1→ 4) D Glu。 相似文献
18.
The synthesis of glycosides by enzymatic transglycosylation is a kinetically controlled reaction performed in the context of a non-favorable thermodynamic equilibrium. An unreactive organic cosolvent which increases the selectivity of the enzyme for glycosyl transfer to the acceptor nucleophile compared with water (Ksel) could improve maximum product yield. Here we report on the effect of the ionic liquid 1,3-dimethylimidazoliummethylsulfate on hydrolase and transferase activities of the hyperthermostable beta-glycosidase CelB from the archaeon Pyrococcus furiosus. CelB retained full catalytic efficiency for lactose hydrolysis at 80 degrees C in a 50% (by vol.) solution of ionic liquid in sodium citrate buffer, pH 5.5. It was inactive but not irreversibly denatured at 70% ionic liquid. Using lactose (0.15 M) as galactosyl donor, values of Ksel for a representative series of eight acceptor alcohols were determined in kinetic assays at 80 degrees C and found to increase between 1.3-fold (D-xylose) and 3.1-fold (glycerol) in 45% ionic liquid. Enhancement of Ksel was dependent on ionic liquid concentration and higher than expected from the decrease in water activity caused by the cosolvent. Experimental molar ratios of D-glucose and D-galactose produced during enzymatic conversion of lactose (75-150 mM) in the presence of D-xylose (0.5 M) or glycerol (0.5 M) showed excellent agreement with predictions based on Ksel values and confirm a significant, yet moderate effect of 45% ionic liquid on increasing the yield of D-galactoside product, by < or = 10%. 相似文献
19.
Lailatul QadariyahMahfud SumarnoSiti Machmudah WahyudionoMitsuru Sasaki Motonobu Goto 《Bioresource technology》2011,102(19):9267-9271
Sub-critical or supercritical water was utilized for the degradation of glycerol in an environmentally benign reaction. The reaction was carried out in a batch reactor in the temperature range of 473-673 K, pressure of 30 MPa, and reaction time of 20-60 min. The effects of temperature and reaction time were observed. The degradation of glycerol produced acetaldehyde, acrolein, allyl alcohol and un-identified products. The highest yield of acrolein, acetaldehyde and allyl alcohol were 0.20, 7.17, 96.69 mol%, respectively. Glycerol conversion was 99.92 mol%. While acetaldehyde was formed only in sub-critical water and allyl alcohol only in supercritical water, acrolein was formed in both. The kinetics of the global reaction displayed a pseudo-first-order. The activation energy at subcritical water was 39.6 kJ/mol. Based on the results, this method could be an efficient method for glycerol degradation because the high conversion of glycerol was obtained. 相似文献
20.
Condensation of Cbz-Asp and PheOMe catalyzed by a neutral protease from Vibrio sp. T1800 (Vimelysin: VLN) was studied. VLN showed a relatively higher catalytic activity of condensation and an apparently larger yield after 3 h or 24 h, in comparison with thermolysin (TLN), especially at lower pH and temperatures.VLN showed higher solvent-tolerance than TLN. TLhe apparent highest yield (25%) was obtained in 30% DMSO by using VLN; under similar conditions, TLN gave only about a half of this value. The rate of the condensation reaction per mole of enzyme (v/[E](o)) in DMSO 50% at 37 degrees C and pH 6.5 was 0.16 s(-1) for VLN and 0.047 s(-1) for TLN. In 30% ethanol VLN showed more than three-fold peptide yield than TLN after 5 h reaction. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 387-390, 1997. 相似文献