红色诺卡氏菌细胞壁骨架辅助治疗肿瘤作用的研究个例分析及文献复习

摘要 目的：探究红色诺卡氏菌细胞壁骨架(Nocardia rubra cell wall skeleton, N-CWS)辅助治疗肿瘤作用的研究。方法：在北京协和医院肿瘤内科近5年收治的实体肿瘤患者中,选取愿意接受N-CWS辅助治疗的患者,皮下注射红色诺卡氏菌细胞壁骨架（商品名艾克佳）,200～400 μg/次,每周2次,监测其肿瘤标志物血清癌胚抗原(serum carcinoembryonic antigen, CEA)、影像学及体力状况评分(eastern cooperative oncology group,ECOG评分)等变化,评估N-CWS疗效。结果：病例1加用N-CWS后CEA下降并保持低值4月左右,ECOG评分保持在0~1分。病例2加用N-CWS后CEA下降并维持稳定2月,ECOG 0~1分,患者自觉使用后体力较前好转。病例3在化疗6程后CEA保持最低,并保持稳定1个月。ECOG 保持在0~2分。病例1和病例2的CEA保持稳定的时间长于病例3,且ECOG评分保持低于病例3。结论：使用N-CWS辅助治疗肿瘤,患者CEA水平下降,体力状况改善,显著的改善了患者的症状。     

Protoplast preparation and regeneration in Nocardia mediterranei

Abstract A standard procedure for the preparation of viable Streptomyces protoplasts was found to be unsuccessful for the rifamycin-producing actinomycete Nocardia mediterranei . An alternative method for N. mediterranei protoplast preparation is described. This method was also found to be applicable to several other ansamycin-producing actinomycetes.     

In vitro augmentation of cytotoxicity by N-CWS in peritoneal polymorphonuclear leukocytes (PMNs) induced by PSK,OK-432 or N-CWS

Peritoneal polymorponuclear leukocytes (PMNs) were collected from the peritoneal cavity of C3H/He mice 6 hrs after intraperitoneal (i.p.) injection of 2.5 mg/head of PSK, 1 KE (100 µg)/head of OK-432 or 200 µg/head ofNocardia rubra cell wall skeleton (N-CWS). Withoutin vitro stimulation, these PMNs did not show cytotoxicity to syngeneic MM46 mammary carcinoma cells in⁵¹Cr release assay. Cytotoxicity of these PMNs was augmented by the addition of 25 µg/ml of N-CWS but not of PSK or OK-432 to cultures for the assay at the beginning of the culture. H₂O₂ production of PSK-induced PMNs was increased by thein vitro addition of 25 µg/ml of N-CWS but not of PSK. These results suggest that PSK as well as OK-432 and N-CWS can induce PMNs capable of responding further to N-CWS as the second stimulant.     

EPR and Mössbauer spectroscopic studies on the tetrameric,NAD-linked hydrogenase of Nocardia opaca 1b and its two dimers: 1. The βδ-dimer—a prototype of a simple hydrogenase

The cytoplasmic, tetrameric NAD-linked hydrogenase from Nocardia opaca 1b can be separated in two dimeric substructures, an -dimer with NADH:electron acceptor oxidoreductase (diaphorase) activity and a -dimer which displays hydrogenase activity with artificial electron carriers. These two dimers were preparatively isolated by a FPLC Mono Q procedure in the absence of nickel and at alkaline pH values. The hydrogenase-active -dimer contained, as analyzed by inductively coupled plasma mass spectrometry (ICP-MS), 3.5–3.9 iron atoms and 1.3–1.7 nickel atoms per dimer molecule. EPR and Mössbauer spectra indicated the presence of a [4Fe-4S] cluster. This center turned out to be extremely labile towards oxidants. Oxidation led to irreversible convertion into a [3Fe-4S] form, thus representing an artifact and not a regulatory state of the cluster. The midpoint redox potential of the [4Fe-4S] cluster was determined to be -385 mV. Very weak EPR Ni signals of the -dimer were detectable in the oxidized as well as in the reduced state. The diaphorase-active -dimer was free of nickel and the iron content corresponded to 11.2–12.8 Fe atoms per dimer molecule. From EPR and Mössbauer measurements it was concluded that this dimer contained two [4Fe-4S] clusters, one [2Fe-2S] and one [3Fe-4S] cluster. In accordance with the results obtained for the dimer proteins, for the whole enzyme an iron content of 15.8–16.2 atoms per enzyme molecule have been determined. EPR spectra and spectrum simulations of the native hydrogenase corroborate the cluster assignments of the two dimers: in total the enzyme contains one [2Fe-2S] cluster, one [3Fe-4S] cluster and three [4Fe-4S] clusters.     

Molecular genetic evidence for the transfer of Oerskovia species into the genus Cellulomonas

A close genetic relationship among strains of Oerskovia turbata, O. xanthineolytica and various coryneforms is indicated by DNA-DNA reassociation studies. O. xanthineolytica shares high homology values (over 60%) with Cellulomonas cartae, Nocardia cellulans, Brevibacterium fermentans and Corynebacterium manihot. O. turbata and other cellulomonads show lower DNA homology values (20–25%) which are still high enough, however, to indicate a relationship at the genus level. Based on these data and supported by comparative analysis of the ribosomal 16 S RNA and the similarity of peptidoglycan types, the transfer of Oerskovia species into the genus Cellulomonas is justified.This paper is respectively dedicated to our teacher and mentor, Professor Dr. O. Kandler, on the occasion of his 60th birthday.     

Epoxidation of propylene utilizing Nocardia corallina immobilized by gel entrapment or adsorption

Nocardia corallina B-276 cells are capable of catalysing the direct epoxidation of propylene to propylene oxide, through a monooxygenase enzyme system. The present work was undertaken to see how immobilization of the whole cells by adsorption or entrapment on solid supports would influence the rate and duration of the epoxidation activity. With immobilization by adsorption, the propylene oxide forming activity was highest on hydrophobic supports, such as polypropylene or polyethylene. Under certain conditions the activity was three times that of the free control cells. However, the duration of epoxidation activity was considerably less for the adsorbed cells. The cell loading by adsorption, determined with¹⁴C-labelled cells was 1.0–2.8 mg dry cell weight g^?1of support. The cells, whether immobilized or not (controls), were tested using a continuous gas flow packed-bed or bubble-type reactor. Immobilization of the cells by entrapment in calcium alginate beads gave about the same propylene oxide forming activity and stability as with control cells, provided the reactor was operated at low temperature (30°C) and low oxygen content (20%) of the feed stream. The results suggest that entrapment in a hydrophobic matrix might be a more favourable system; although additional investigation of the rate limiting steps as well as the cause of the activity loss with time is needed.     

用诺卡氏菌酶法转化顺式环氧琥珀酸生产L（＋）—酒石酸的研究

L(+)酒石酸(又称d-酒石酸),是一种重要的食品添加剂和医药拆分剂,在食品工业、医药工业和化学工业中有广泛用途。传统的L(+)酒石酸是从葡萄酒副产物酒石中提取的。世界主要厂商在西欧。随着世界经济发展,对酒石酸的需求量越来越大,由于酒石供应量有限,酒石酸因此不能满足供应。而且常常受气候、葡萄种植业、葡萄酒酿造业的影响,导致较大的价格波动。因此开发新的酒石酸生产方法就必然引起人们重视。 早年(1929年)就有发酵法直接从葡萄     

Conversion of 3-cyanopyridine to nicotinic acid by Nocardia rhodochrous LL100-21

3-Cyanopyridinase activity, i.e. the ability to convert 3-cyanopyridine to nicotinic acid plus ammonia, was induced in stationary phase cultures of Nocardia rhodochrous LL100-21 by the addition of 2-, 3-, or 4-cyanopyridine or benzonitrile; the latter nitrile gave maximum induction. Harvested bacteria possessing 3-cyanopyridinase activity could stoichiometrically convert 3-cyanopyridine at concentrations of up to 0.5 to nicotinic acid. Both 3-cyanopyridine and nicotinic acid inhibited the hydrolysis of 3-cyanopyridine by intact bacteria. Bacteria immobilized in calcium alginate beads and used in column bioreactors retained 3-cyanopyridinase activity for over 150 h when continuously supplied with 0.3 3-cyanopyridine.     

The biosynthetic origin of the pyridone ring of efrotomycin

Summary Nocardialactamdurans has been shown to catabolyse uracil via the reductive pathway. The end product of this pathway, -alanine, is incorporated into the pyridone ring of efrotomycin. Support for this proposal includes: (1) reversal of thymine inhibition of efrotomycin biosynthesis by dihydrouracil andN-carbamoyl--aline, two intermediates of the catabolic pathway; (2) incorporation of [5,6-³H]-uracil into efrotomycin with a relative molar specific activity of approximately 0.5, close to the theoretical maximum; and (3)¹³C coupling at C₄ and C₅ of efrotomycin after feeding resting cells with [4,5-¹³C]-uracil. Our results do not rule out the possibility of an alternative source of -alanine or the coexistence of uracil catabolism via oxidative reactions.