Expression of functional Candida antarctica lipase B in a cell‐free protein synthesis system derived from Escherichia coli

This article reports the cell‐free expression of functional Lipase B from Candida antarctica (CalB) in an Escherichia coli extract. Although most of the cell‐free synthesized CalB was insoluble under conventional reaction conditions, the combined use of molecular chaperones led to the soluble expression of CalB. In addition, the functional enzyme was generated by applying the optimal redox potential. When examined using p‐nitrophenyl palmitate as a substrate, the specific activity of the cell‐free synthesized CalB was higher than that of the reference protein produced in Pichia pastoris. These results highlight the potential of cell‐free protein synthesis technology as a powerful platform for the rapid expression, screening and analysis of industrially important enzymes. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Saponins from Lysimachia candida

Three saponins were isolated from Lysimachia candida Lindl. By spectral and chemical analysis, they were identified as primulagenin A-3-O-β-D-xylopyranosy( 1→2)-β-D-glucopyranosyl( 1→4)-[β-D-glu- copyranosyl(1→2) ] -a-L-arabinopyranoside ( 1 ), protoprimulagenin A-3-O-β-D-xylopyranosyl( 1→2)-β-D-glu- copyranosyl( 1→4)-[ β-D-glucopyranosyl ( 1 →2) ]-a-L-arabinopyranoside ( lysikoianoside, 2) and a-spinas- teml-glucopyranoside (3) respectively. 1 is a new compound named candidoside.     

单条草的皂甙成分

从单条草（ＬｙｓｉｍａｃｈｉａｃａｎｄｉｄａＬｉｎｄｌ）全草中分离出３个皂甙类化合物,经波谱分析并结合化学方法鉴定为：报春花素Ａ３ＯβＤ吡喃木糖基（１→２）βＤ吡喃葡萄糖基（１→４）［βＤ吡喃葡萄糖基（１→２）］αＬ吡喃阿拉伯糖甙（１）、原报春花素Ａ３ＯβＤ吡喃木糖基（１→２）βＤ吡喃葡萄糖基（１→４）［βＤ吡喃葡萄糖基（１→２）］αＬ吡喃阿拉伯糖甙（ｌｙｓｉｋｏｉａｎｏｓｉｄｅ,２）和α菠甾醇葡萄糖甙（３）。其中１是新化合物,命名为单条草甙（ｃａｎｄｉｄｏｓｉｄｅ）。     

Production of Tumor Necrosis Factor-α in Granulocytopenic Mice with Pulmonary Candidiasis and Its Modification with Granulocyte Colony-Stimulating Factor

In the present study, a lethal model of pulmonary candidiasis was established using granulocytopenic mice with cyclophosphamide. These mice started to die 1 day after infection and had all died within the next 48 hr. The counts of live C. albicans in the lung gradually increased with time, while the organisms were quickly eliminated in the normal mice. From the histology and measurements on bronchoalveolar lavage fluid (BALF), polymorphonuclear cells (PMN) response was almost zero up to 24 hr, and then a weak but significant response was observed at 48 hr, while a marked accumulation of PMN was detected from as early as 6 hr in normal mice. In contrast, macrophages had accumulated in BALF by 48 hr in granulocytopenic mice, but not in normal mice. Both in serum and BALF, a considerable level of tumor necrosis factor-α (TNF-α) was detected from 6 hr, peaking at 24 to 48 hr, while in normal mice the quantity was under the detection limit in serum and very low in BALF. The effects of administering granulocyte colony-stimulating factor (G-CSF) on these parameters were next examined. G-CSF significantly prolonged the survival time of granulocytopenic mice, promoted the clearance of organisms through increasing the counts of PMN in the lung, and strongly inhibited the production of TNF-α both in BALF and serum. These results suggest that this cytokine does not protect them, but plays some role in their death due to candidial infection in granulocytopenic mice.     

Specific microbial colonizations in the periodontal sites of HIV-infected subjects.

The purpose of this study was to examine colonization by specific organisms at periodontal sites in HIV-seropositive [HIV(+)] subjects. A total of 67 HIV(+) and 32 HIV(--) subjects were investigated. The specific pathogens included black-pigmented anaerobic rods (BPAR), Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Candida albicans and mycoplasma species. P. gingivalis was present in the HIV(+) subjects more frequently than in the HIV(--) periodontitis patients (P < 0.01). The cell numbers of BPAR and P. gingivalis and percentages as the total of CFUs on blood agar cultured in an anaerobic chamber were statistically higher in periodontal pocket samples from HIV(+) than from HIV(--). A. actinomycetemcomitans was also detected at a high rate (41.8%) in HIV(+) patients. The average cell numbers of C. albicans were higher in samples from the HIV(+) group (P < 0.05). The detection rate of mycoplasma species in the HIV(+) patients was significantly lower than that in HIV(--) subjects (P < 0.05), and most isolated mycoplasma strains were Mycoplasma salivarium.     

Prophylactic Effect of Enterococcus faecalis FK-23 Preparation on Experimental Candidiasis in Mice

The prophylactic effects of heat-killed cells of Enterococcus faecalis FK-23 (FK-23 preparation) on experimental candidiasis were investigated in normal and leukopenic mice. In cyclophosphamide-induced leukopenic mice, oral or intraperitoneal administration of the FK-23 preparation at a daily dose of 1.25 or 5 mg/mouse for 3 consecutive days prior to Candida albicans infection significantly prolonged survival periods of the infected mice, and decreased viable counts of C. albicans recovered from their kidneys. In normal mice, the FK-23 preparation administered at dosages ranging from 0.63 to 10 mg/mouse/day for 3 consecutive days was ineffective, while in leukopenic mice, the FK-23 administered orally caused a facilitated recovery in the number of white blood cells including neutrophils. Furthermore, intraperitoneal administration of the FK-23 preparation into mice augmented the anti-Candida activity of immunocompromised peritoneal exudate cells obtained from the animals. These results suggested the potential usefulness of the FK-23 preparation as a prophylactic agent for the management of patients with opportunistic fungal infections.     

Nonconstricting-ring formation in two species of nematode-capturing hyphomycetes

 Dactylella leptospora and Dactylaria candida, and Arthrobotrys dactyloides (Hyphomycetes), capture nematodes by nonconstricting- and constricting-ring traps, respectively. In the formation of the constricting-ring trap of the latter fungus, the basal portion of a curved hyphal branch put forth a bud to fuse with its advancing tip to make a ring. However, in nonconstricting-ring formation in the former two fungi, the portion behind the tip of the curved branch did not develop such a bud before fusion with the tip. Received: May 31, 2002 / Accepted: July 5, 2002 Correspondence to:M. Saikawa