维生素B12发酵工艺进展

综述了维生素B12(VB12)发酵工艺路线、菌种选育、工艺优化等方面的研究进展,展望了VB12发酵工艺发展前景。     

UBF工艺处理维生素B_（12）生产废水的研究

维生素B12生产废水属高CODCr、高氨氮废水,但B/C值达到0.4以上,且原辅料中抑制性物质较少,可生化性较好,适宜采用生物工艺进行处理。采用厌氧技术既可节省运行费用,又可产生沼气,增加效益;UBF是新型复合式厌氧流化床反应器,能够有效处理高浓度有机废水,通过实验可以确定使用UBF工艺处理维生素B12生产过程中产生废水的可行性,为废水处理提供了技术依据。实验结果表明,UBF工艺处理维生素B12生产废水,在使用絮状污泥的情况下,CODCr去除率可达到85%,进水容积负荷达到5 kg/m3·d,是一条可行的工艺路线。     

Solution Structure and Constrained Molecular Dynamics Study of Vitamin B12 Conjugates of the Anorectic Peptide PYY(3–36)

Vitamin B₁₂–peptide conjugates have considerable therapeutic potential through improved pharmacokinetic and/or pharmacodynamic properties imparted on the peptide upon covalent attachment to vitamin B₁₂ (B₁₂). There remains a lack of structural studies investigating the effects of B₁₂ conjugation on peptide secondary structure. Determining the solution structure of a B₁₂–peptide conjugate or conjugates and measuring functions of the conjugate(s) at the target peptide receptor may offer considerable insight concerning the future design of fully optimized conjugates. This methodology is especially useful in tandem with constrained molecular dynamics (MD) studies, such that predictions may be made about conjugates not yet synthesized. Focusing on two B₁₂ conjugates of the anorectic peptide PYY(3–36), one of which was previously demonstrated to have improved food intake reduction compared with PYY(3–36), we performed NMR structural analyses and used the information to conduct MD simulations. The study provides rare structural insight into vitamin B₁₂ conjugates and validates the fact that B₁₂ can be conjugated to a peptide without markedly affecting peptide secondary structure.     

维生素B12的技术进展

维生素B12是人和动物体内霞要的水溶性维生素之一。综述了维生素B12的性质、合成及生产情况。工业生产中,脱氮假单胞杆菌和费氏丙酸杆菌是主要的生产菌种,有厌氧和好氧2种生产工艺。今后应培养高产菌株,改进发酵工艺。     

Release of vitamin B12 from poly(N‐vinyl‐2‐pyrrolidone)‐crosslinked polyacrylamide hydrogels: A kinetic study

Hydrogels, composed of poly(N‐vinyl‐2‐pyrrolidone) and crosslinked polyacrylamide, were synthesized and the release of vitamin B₁₂ from these hydrogels was studied as a function of the degree of crosslinking and pH of the external swelling media. The three drug‐loaded hydrogel samples synthesized with different crosslinking ratios of 0.3, 0.7, and 1.2 (in mol %) follow different drug‐release mechanisms, that is, chain relaxation with zero‐order, non‐Fickian and Fickian, or diffusion‐controlled mechanisms. To establish a correlation between their swelling behavior and drug‐release mechanism, the former was studied by the weight‐gain method and, at the same time, the concentration of the drug released was studied colorimetrically. Various swelling parameters such as the swelling exponent n, gel‐characteristic constant k, penetration velocity v, and diffusion coefficient D were evaluated to reflect the quantitative aspect of the swelling behavior of these hydrogels. Finally, the drug‐release behavior of the hydrogels was explained by proposing the swelling‐dependent mechanism. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 1706–1714, 2000     

Study on the dissolved oxygen control strategy in large‐scale vitamin B12 fermentation by Pseudomonas denitrificans

BACKGROUND: There are two different routes for vitamin B₁₂ biosynthesis, which results in discrepancies and uncertainties of the dissolved oxygen (DO) concentration for vitamin B₁₂ fermentation. In this paper, the DO control strategy was explored for industrial vitamin B₁₂ fermentation by Pesudomonas denitrificans in 120000‐L fermenter. RESULTS: A DO‐stat strategy was first successfully scaled up from a 9000 L fermenter to a 120 000 L fermenter. Then a multi‐stage DO control strategy was further established in the 120 000 L fermenter, in which the DO level was shifted from 8–10% (20–48 h) to 2–5% (49–106 h) and below 2% (107–168 h) by gradually reducing the rate of aeration and agitation. As a result, 198.80 mg L^?1 of vitamin B₁₂ was obtained, which was significantly higher than those obtained under the fermentations with one‐stage DO control. CONCLUSIONS: The comparatively low DO level was favorable for vitamin B₁₂ biosynthesis, but it would have an extremely negative effect on cell growth. Compared with the low DO level maintained at all times of the fermentation process, a multi‐stage DO control strategy could not only increase the biomass but also improve vitamin B₁₂ biosynthesis. Copyright © 2012 Society of Chemical Industry     

Phenoxaphosphino‐Modified Xantphos‐Type Ligands in the Rhodium‐Catalysed Hydroformylation of Internal and Terminal Alkenes

The solubility of the modifying ligand is an important parameter for the efficiency of a rhodium‐catalysed hydroformylation system. A facile synthetic procedure for the preparation of well‐defined xanthene‐type ligands was developed in order to study the influence of alkyl substituents at the 2‐, and 7‐positions of the 9,9‐dimethylxanthene backbone and at the 2‐, and 8‐positions of the phenoxaphosphino moiety of ligands 1 – 16 on solubility in toluene and the influence of these substituents on the performance of the ligands in the rhodium‐catalysed hydroformylation. An increase in solubility from 2.3 mmol⋅L⁻¹ to >495 mmol⋅L⁻¹ was observed from the least soluble to the most soluble ligand. A solubility of at least 58 mmol⋅L⁻¹ was estimated to be sufficient for a large‐scale application of these ligands in hydroformylation. Highly active and selective catalysts for the rhodium‐catalysed hydroformylation of 1‐octene and trans‐2‐octene to nonanal, and for the hydroformylation of 2‐pentene to hexanal were obtained by employing these ligands. Average rates of >1600 (mol aldehyde) × (mol Rh)⁻¹×h⁻¹ {conditions: p(CO/H₂) = 20 bar, T = 353 K, [Rh] = 1 mM, [alkene] = 637 mM} and excellent regio‐selectivities of up to 99% toward the linear product were obtained when 1‐octene was used as substrate. For internal olefins average rates of >145 (mol aldehyde)×(mol Rh)⁻¹×h⁻¹ {p(CO/H₂) = 3.6–10 bar, T = 393 K, [Rh] = 1 mM, [alkene] = 640–928 mM} and high regio‐selectivities up to 91% toward the linear product were obtained.     

超滤膜分离技术在维生素B12生产中的应用

采用U1tra—flo超滤系统去除不经预处理的维生素B12发酵液中残留的菌丝体和蛋白质等杂质。用截留分了量为10万的UItra—flo膜过滤维生素B12发酵液,滤液质量高,平均膜通量可达到122．52L／m2·h^-1,膜通量衰减较慢,被污染的膜经清洗后与新膜没有明显的差异,说明Ultra-flo超滤系统处理能力强且可长时间稳定运行。该超滤系统现已在华北制药集团威可达公司成功应用。     

庆大霉素普鲁卡因维B12滴丸的处方工艺研究及质量控制

对庆大霉素普鲁卡因维B12胶囊进行工艺改进,采用固体分散技术制成滴丸剂,提高溶出度。通过对原料粒径、滴丸基质及冷凝剂等进行试验研究,优选出处方及制备工艺,并通过方法学研究,建立起质量控制方法,试验结果表明盐酸普鲁卡因在6.15～14.35μg/mL具有良好的线性关系（y=0.062x＋0.013,R2=0.999）,该方法重复性好,准确性高。     

Nonisothermal and isothermal crystallization kinetics of nylon‐12

The isothermal and nonisothermal crystallization behavior of Nylon 12 was investigated using differential scanning calorimetry (DSC). An Avrami analysis was used to study the isothermal crystallization kinetics of Nylon 12, the Avrami exponent (n) determined and its relevance to crystal growth discussed and an activation energy for the process evaluated using an Arrhenius type expression. The Lauritzen and Hoffman analysis was used to examine the spherulitic growth process of the primary crystallization stage of Nylon 12. The surface‐free energy and work of chain folding were calculated using a procedure reported by Hoffmann and the work of chain folding per molecular fold (σ) and chain stiffness of Nylon 12 (q) was calculated and compared to values reported for Nylons 6,6 and 11. The Jeziorny modification of the Avrami analysis, Cazé and Chuah average Avrami parameter methods and Ozawa equation were used in an attempt to model the nonisothermal crystallization kinetics of Nylon 12. A combined Avrami and Ozawa treatment, described by Liu, was used to more accurately model the nonisothermal crystallization kinetics of Nylon 12. The activation energy for nonisothermal crystallization processes was determined using the Kissinger method for Nylon 12 and compared with values reported previously for Nylon 6,6 and Nylon 11. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

纳滤膜技术在维生素B12生产中的应用

为提高维生素B12提取过程中的收率、降低水资源消耗,采用纳滤膜对维生素B12超滤滤液进行浓缩。考察了pH值、压力、浓缩倍数对膜通量的影响。     

The Role of Vitamin D and Vitamin D Binding Protein in Chronic Liver Diseases

Vitamin D (calciferol) is a fat-soluble vitamin that has a significant role in phospho-calcium metabolism, maintaining normal calcium levels and bone health development. The most important compounds of vitamin D are cholecalciferol (vitamin D3, or VD3) and ergocalciferol (vitamin D2, or VD2). Besides its major role in maintaining an adequate level of calcium and phosphate concentrations, vitamin D is involved in cell growth and differentiation and immune function. Recently, the association between vitamin D deficiency and the progression of fibrosis in chronic liver disease (CLD) was confirmed, given the hepatic activation process and high prevalence of vitamin D deficiency in these diseases. There are reports of vitamin D deficiency in CLD regardless of the etiology (chronic viral hepatitis, alcoholic cirrhosis, non-alcoholic fatty liver disease, primary biliary cirrhosis, or autoimmune hepatitis). Vitamin D binding protein (VDBP) is synthesized by the liver and has the role of binding and transporting vitamin D and its metabolites to the target organs. VDBP also plays an important role in inflammatory response secondary to tissue damage, being involved in the degradation of actin. As intense research during the last decades revealed the possible role of vitamin D in liver diseases, a deeper understanding of the vitamin D, vitamin D receptors (VDRs), and VDBP involvement in liver inflammation and fibrogenesis could represent the basis for the development of new strategies for diagnosis, prognosis, and treatment of liver diseases. This narrative review presents an overview of the evidence of the role of vitamin D and VDBP in CLD, both at the experimental and clinical levels.     

Intrinsic factor and vitamin B12 complex‐loaded poly[lactic‐co‐(glycolic acid)] microspheres: preparation,characterization and drug release

BACKGROUND: Vitamin B12 is an essential vitamin required by all mammals. Absorption of vitamin B12 is facilitated by binding of intrinsic factor–vitamin B₁₂ complex to specific receptors in the ileum. In humans a deficiency of this vitamin or a lack of intrinsic factor leads to pernicious anaemia. The major objective of the present study was to prepare intrinsic factor–vitamin B12 complex‐loaded poly[lactic‐co‐(glycolic acid)] (PLGA)‐based microparticles and to investigate their release kinetics. RESULTS: PLGA copolymer was synthesized by the ring‐opening polymerization method and characterized using gel permeation chromatography, Fourier transform infrared spectroscopy and ¹H NMR. The glass transition temperature measurement showed a single T_g at 40 °C. The intrinsic factor–vitamin B12 complex‐loaded PLGA microspheres were prepared by a water‐in‐oil‐in‐water double emulsion solvent extraction/evaporation technique. An environmental scanning electron microscopy investigation demonstrated that the PLGA particles had a mean particle diameter of 38 µm. Interestingly, different drug release patterns (bi‐ and triphasic ones) were observed for vitamin B12‐loaded and intrinsic factor–vitamin B12 complex‐loaded microspheres. In contrast to the rapid release of vitamin B12 by itself, in vitro release tests showed that intrinsic factor and vitamin B12 in the complex were released from PLGA microspheres in a sustained manner over 15 days. CONCLUSION: PLGA microspheres can be an effective carrier for the intrinsic factor–vitamin B12 complex. Copyright © 2007 Society of Chemical Industry     

Ferrocenyl‐Aryl Based trans‐Chelating Diphosphine Ligands: Synthesis,Molecular Structure and Application in Enantioselective Hydrogenation

Potentially trans‐chelating diphosphine ligands based on a ferrocenyl‐aryl backbone were synthesised in a four‐step sequence and the molecular structures in the solid state of two representatives were determined by X‐ray diffraction. High throughput screening of these ligands in rhodium‐, ruthenium‐ and iridium‐mediated hydrogenations of a variety of alkenes and ketones revealed that these ligands can deliver high enantioselectivity for alkenes (up to 98% ee) but are less selective when ketones are used as the substrates. The coordination behaviour of one ligand in its square planar palladium and platinum dichloride complexes was studied by ³¹P NMR and only trans‐chelated complexes, together with oligomeric by‐products, were observed. Reaction with the (p‐cymene)ruthenium dichloride dimer, [RuCl₂(p‐cymene)]₂, resulted in a mixture of diastereomeric complexes.     

细胞膜分离费氏丙酸杆菌离位转化合成维生素B12

采用膜分离技术将费氏丙酸杆菌菌体细胞分离,离位合成转化维生素B12,考察了不同类型及孔径的膜对发酵液的处理能力及对菌体细胞活性的损伤情况,分析了不同浓缩条件下维生素B12的转化效果. 结果表明,卷式膜比陶瓷膜更适用于发酵液中菌体细胞的分离,用0.22 mm卷式膜处理时,细胞的生物活性几乎不受损伤,存活率达99.5%,将发酵液浓缩4倍,中间体Abo-cbi转化率达78.8%,维生素B12产量为53.9 mg/L.     

石墨烯量子点－VB2荧光能量转移法测定 VB2

以氧化石墨烯为单一碳源,通过简单的超声－水热法制备了形貌均一、单分散性好、尺寸约为5 nm的石墨烯量子点（ GQD）。所制备的GQD不仅具有强的蓝色荧光发射,而且具有激发波长依赖性荧光。以GQD作为能量供体,以维生素B2（ VB2）为能量受体,建立了一种荧光能量转移体系检测VB2含量的方法。在优化条件下,本方法的检测范围为1．0×10^－6～3．0×10^－5 mol· L^－1,相关系数为0．9978,检出限为3．3×10^－7 mol· L^－1。