Abstract: Apparatuses and methods for providing thermal pathways from a substrate to a thermal bonding pad. The thermal pathways may be metal extensions of the thermal bonding pad that are disposed in channels formed in a backside passivation layer underneath the thermal bonding pad, and may be in direct contact with an underlying substrate. The thermal pathways may provide improved thermal dissipation from the substrate.

Abstract: A method for wafer level packaging includes forming one or more die, forming a plated metal ring (PMR) on each die, forming a cover wafer (CW), the CW having one or more plated seal rings, forming a body wafer (BW), the BW having cavities and a metal layer on a first side of the BW, aligning a respective die to the CW so that a PMR on the respective die is aligned to a respective plated seal ring (PSR) on the CW, bonding the PMR on the respective die to the respective PSR, aligning the BW to the CW so that a respective cavity of the BW surrounds each respective die bonded to the CW and so that the metal layer on the BW is aligned with at least one PSR on the CW, and bonding the metal layer on the first side of the BW to the PSR on the CW. Each PMR has a first height and each PSR has a second height.

Abstract: Biogas is obtained in a solid-state anaerobic digester from a solid organic biomass and a digestion effluent produced in an associated liquid anaerobic digester. The solid organic biomass and digestion effluent are mixed to produce an effluent-biomass mixture. The effluent-biomass mixture is then incubated in the solid state anaerobic digester, producing a biogas and a digestate. The incubation is controlled by adjusting the composition and properties of the effluent-biomass mixture. The solid organic biomass may comprise lignocellulosic biomass, food waste, agricultural waste and the like.

Abstract: The present invention relates to a method of manufacturing a cloverleaf microgyroscope containing an integrated post comprising: attaching a post wafer to a resonator wafer, forming a bottom post from the post wafer being attached to the resonator wafer, preparing a base wafer with through-wafer interconnects, attaching the resonator wafer to the base wafer, wherein the bottom post fits into a post hole in the base wafer, forming a top post from the resonator wafer, wherein the bottom and top post are formed symmetrically around the same axis, and attaching a cap wafer on top of the base wafer.

Abstract: An isolated DNA sequence that encodes a peptide with CPC-acetylhydrolase enzymatic activity is disclosed. Also, a method of expressing CPC-acetylhydrolase activity that includes the steps of (a) providing a microorganism that is susceptible to transformation with the isolated DNA sequence and that, upon transformation, expresses the CPC-acetylhydrolase activity encoded by the sequence, and (b) transforming the microorganism with the DNA sequence to cause expression of the CPC-acetylhydrolase activity in the microorganism is disclosed. Also, a method for producing a microorganism with increased capacity to aid in the production of cephalosporin, which method includes the steps of (a) providing a microorganism that has CPC-AH activity by virtue of expression of the DNA sequence or of a fragment thereof coding for the CPC-acetylhydrolase enzyme activity; and (b) inactivating the activity by disrupting expression of the DNA sequence is disclosed.

Abstract: An enzymatic process for preparing 3-thiolated 7-aminocephalosporanic acid derivatives comprises the steps of enzymatically converting a 3-thiolated cephalosporin C of the formula I: 1

Abstract: An enzyme in a mixture containing the enzyme and a contaminant enzyme is purified by selectively aggregating and precipitating the contaminant enzyme with a surfactant. A deacetylase contaminant is separated from Cephalosporin C acylase using a cationic surfactant in an amount of 0.1 to 0.6%. Surfactants include benzylated or methylated cationic surfactants such as alkyl(palm)dimethylbenzyl ammonium chloride, alkyl(palm)trimethyl ammonium chloride and dodecyltrimethyl ammonium chloride. An immobilized enzyme is regenerated using a protease to remove the enzyme from a carrier. The carrier may be a synthetic adsorbent or an ion exchange resin having pores of 100 nm or less in diameter, and the immobilized enzyme is optionally crosslinked. Immobilized cephalosporin C acylase is regenerated using an alkaline or acidic protease.

Abstract: The present invention discloses a process for the production of N-deacylated cephalosporin compounds via the fermentative production of their 7-acylated counterparts.

Abstract: The invention relates to a process for the preparation of an antibiotic, in particular cefalexin, ampicillin, amoxicillin, cefaclor, cefradin, cefadroxil, cefotaxim and the like, wherein a beta-lactam core is acylated, the antibiotic is recovered from the reaction mixture, the remaining mother liquor is subjected to a hydrolysis reaction in which the antibiotic present in the mother liquor is decomposed into its initial compounds, in particular the beta-lactam core, and the acylation agent is hydrolized. The beta-lactam core can then be recovered virtually quantitatively or recycled to the acylation reaction. This is because it has been found that the solubility of the beta-lactam core is unexpectedly high at relatively high concentrations of acylation agent and antibiotic.

Abstract: This invention provides a waste-processing system capable of processing high-solids wastes such as manure. This invention provides a compact U-shaped digester that allows for recycling of activated sludge to improve the efficiency of the process. Efficiency is also improved through a sludge heating design that creates a current in the digester and efficiently heats the sludge. A composter is provided to further process the sludge through aerobic digestion to create usable fertilizer. Finally, one embodiment provides a turbine that is fueled by biological gases from the digester to generate heat and electricity to be used by the system.

Abstract: The invention provides a glutaryl 7-ACA amidase from Pseudomonas diminuta strain BS-203, which catalyzes the hydrolysis of 7-&bgr;-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.

Abstract: The present invention relates to a nucleic acid molecule which codes for the cephalosporin acetylesterase from Bacillus subtilis ATCC 6633 (DSM 11909), vectors and host cells which comprise such a nucleic acid molecule, a process for the recombinant preparation of cephalosporin acetylesterase from B. subtilis ATCC 6633 (DSM 1 1909) using said nucleic acid molecule, and a process for preparing 3-deacetylcephalosporin compounds.

Abstract: A process for the preparation of Cefuroxime acid (I), which comprises the following steps: (1) Extraction of deacetyl 7-glutaryl ACA (II) aqueous solution at acid pH with organic solvents (for example according to the procedures disclosed in U.S. Pat. No. 5,801,241); (2) drying the resulting solution while preventing lactonization of the intermediate; (3) carbamoylation of the hydroxymethyl group at the 3-position by reaction with chlorosulfonyl isocyanate or similar products; (7) extraction of the carbamoyl derivative from step 3 with water at neutral pH; (8) enzymatic hydrolysis of the amide at the 7-position of the cephalosporanic ring with glutaryl acylase; (6) acylation of the amnino group by condensation with 2-furanyl(sin-methoxyimino)acetic acid chloride or mixed anhydride.

Abstract: The present invention disclosures a process for the production of N-deacylated cephalosporin compounds via the fermentative production of their 7-acylated counterparts.

Abstract: A first compound is converted to a second compound enzymatically in a solvent mixture containing water and a fluorinated, non-chlorinated alkane, alkene, or alkyne having up to 4 carbon atoms. Lactams, for example 6-aminopenicillanic acid (6-APA), may be prepared by enzymatic conversion of a first compound, for example penicillin-G, in a solvent mixture comprising water and a non-aqueous organic solvent, for example 1,1,1, 2-tetrafluoroethane. The 6-APA can be caused to precipitate, isolated by filtration and optionally derivatized to produce a desired compound. A by-product of the enzymatic conversion, for example phenylacetic acid, can be isolated by solvent extraction, suitably using a solvent which also comprises 1, 1, 1, 2-tetrafluoroethane.

Abstract: The present invention relates to a novel process for the preparation of cephalosporins having been deacylated at the 7-amino group, by fermentation of a cephalosporin producing microorganism in the presence of a side chain precursor, extraction of the N-substituted cephalosporin compound as present in the fermentation broth or fluid to an organic solvent, back extraction of the N-substituted cephalosporin compound to water, treatment of the aqueous phase with a dicarboxylate acylase and isolation of the crystalline cephalosporin compound according to formula (1) from the conversion solution, characterized in that the fermentation broth or fluid is incubated at acidic conditions and an elevated temperature prior to extraction of the N-substituted cephalosporin compound to an organic solvent.

Abstract: An enzyme in a mixture containing the enzyme and a contaminant enzyme is purified by selectively aggregating and precipitating the contaminant enzyme with a surfactant. A carrier containing an immobilized enzyme is regenerated by using a protease to remove the enzyme from the carrier. Cephalosporin C acylase is purified from a mixture of the acylase and a deacetylase contaminant by selectively aggregating and precipitating the deacetylase contaminant by adding to the solution a benzylated cationic or methylated cationic surfactant in an amount of 0.1 to 0.6% of the mixture. The surfactant may be alkyl(palm)dimethylbenzyl ammonium chloride. A carrier containing immobilized cephalosporin C acylase is regenerated by contacting the carrier with an alkaline or acidic protease. The acylase is immobilized by being bound to the carrier and optionally crosslinked. The carrier may have pores of 100 nm or less in diameter.

Abstract: A bioprocess for preparing adipoyl-7-ACA comprising the steps: (a) transforming cells of a strain of Penicillium chrysogenum which produces isopenicillin N with an expression vector containing DNA encoding an enzyme, having expandase activity capable of accepting adipoyl 6-APA as a substrate, an enzyme having hydroxylase activity capable of accepting adipoyl-7-ADCA as a substrate and an enzyme having acetyl transferase activity capable of accepting adipoyl 7-ADAC as a substrate; (b) culturing the transformed cells from step a) in a suitable culture medium containing an adipate feedstock, wherein said cells produce adipoyl 6-APA; and (c) culturing the transformed cells producing adipoyl 6-APA of step b) under conditions suitable for expression of said DNA encoding enzyme, thereby producing the end product adipoyl-7-ACA.

Abstract: A process is taught for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via enzymatic ring expansion activity on penicillin G, using a Penicillium chrysogenum transformant strain expressing expandase.

Abstract: A process for making 7-aminodeacetyl-cephalosporanic acid (7-ADAC).

Abstract: A mutant CC acylase wherein at least one amino acid at the Ala.sup.49, Met.sup.164, Ser.sup.166, Met.sup.174, Glu.sup.358, Met.sup.465, Met.sup.506, or Met.sup.750 position of the amino acid sequence of the native CC acylase is replaced by a different amino acid, a DNA coding therefor, an expression vector containing the said DNA, a microorganism transformed with the said expression vector, the production of the CC acylase by culturing the said transformant, and use thereof for the production of a compound. The mutant CC acylase of the invention has desirable properties in terms of enzymatic potency, alteration of pH profile, efficiency of processing, and the like.

Abstract: An overall efficient process for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via 3-(carboxyethylthio)propionyl-7-ADCA, using a Penicillium chrysogenum transformant strain expressing expandase in conjunction with acyltransferase, is provided.

Abstract: An enzyme selected from penicillin-G amidase, glutaryl-7-ACA acylase and D-amino acid oxidase is immobilized by covalent bonding on an aminofunctional organosiloxane polymer carrier to provide an immobilized enzyme having a specific volume activity of at least 100 U/g wet carrier. Preferably, the carrier has an average diameter or 0.01 to 3 mm and is essentially spherical. Covalent bonding is accomplished by activating amino groups on the carrier with a dialdehyde and reacting the activated groups with reactive groups on the enzyme. An amount of enzyme is immobilized to provide a weight ratio of enzyme to carrier of 1 to 300 mg protein per g wet carrier.

Abstract: A time serial signal from a signal source is input into a delay circuit, and the signal is read out from arbitrary intermediate taps of the delay circuit to obtain a specific delayed signal. Alternatively, a time serial signal from a signal source is allotted to a plurality of memory circuits, and the allotted signal contents are read at a specific moment to obtain a specific delayed signal. By distributing the delayed signals in this manner, a signal distribution apparatus can send a serial signal, from its beginning with little delay, to multiple terminals requesting the signal at arbitrary times.

Abstract: A method for providing a semisynthetic .beta.-lactam antibiotic by enzyme catalyzed acylation of the parent .beta.-lactam with an activated derivative of the side chain acid wherein a modulator, which consists of one or more compounds different from the reactants and the reaction product and which suppresses the hydrolysis of the activated derivative of the side chain acid and the desired product more than it suppresses the synthesis of the desired product, is added to the reaction mixture, at the beginning of the reaction process, in a concentration from about 0.2 to 100.times.10.sup.3 .mu.m.

Abstract: An overall process for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via enzymatic ring expansion activity on penicillin G, using a Penicillium chrysogenum transformant strain expressing expandase.

Abstract: An overall efficient process for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via 2-(carboxyethylthio)acetyl- and 3-(carboxymethylthio)propionyl-7-ADCA, using a Penicillium chrysogenum transformant strain expressing expandase in conjunction with acyltransferase, is provided.

Abstract: The present invention provides a process for producing 7-aminocephem compounds or salts thereof.

Abstract: Pharmaceutical compositions of an erythromycin derivative combined with sulfisoxazole according to the structural formulas: ##STR1## where R is hydrogen, C1-C10 alkylcarbonyl or C1-C10 alkyl wherein the substituent is amino or cyano; R1 and R2 are independently hydrogen, hydroxyl or amino; and the pharmaceutical salts and esters thereof.

Abstract: Important intermediates for preparing cephalosporin antibiotics, 7-amino-cephalosporanic acid (7-ACA) and 7-aminodeacetylcephalosporanic acid (7-ADAC), are prepared by a novel bioprocess in which a transformed Penicillium chrysogenum strain is cultured in the presence of an adipate feedstock to produce adipoyl-6-APA (6-amino penicillanic acid); followed by the in situ expression of the following genes with which the P. chrysogenum has been transformed:1) an expandase gene, whose expression product converts the adipoyl-6-APA by ring expansion to adipoyl-7-ADCA;2) an hydroxylase gene whose expression product converts the 3-methyl side chain of adipoyl-7-ADCA to 3-hydroxymethyl, to give the first product, 7-aminodeacetylcephalosporanic acid (7-ADAC); and3) an acetyltransferase gene whose expression product converts the 3-hydroxymethyl side chain to the 3-acetyloxymethyl side chain of 7-ACA. The final product, 7-ACA, is then prepared by cleavage of the adipoyl side chain using an adipoyl acylase.

Abstract: Important intermediates for preparing cephalosporin antibiotics, 7-amino-cephalosporanic acid (7-ACA) and 7-aminodeacetylcephalosporanic acid (7-ADAC), are prepared by a novel bioprocess in which a transformed Penicillium chrysogenum strain is cultured in the presence of an adipate feedstock to produce adipoyl-6-APA (6-amino penicillanic acid); followed by the in situ expression of the following genes with which the P. chrysogenum has been transformed:1) an expandase gene, e.g., from Cephalosporium acremonium, whose expression product converts the adipoyl-6-APA by ring expansion to adipoyl-7-ADCA;2) an hydroxylase gene whose expression product converts the 3-methyl side chain of adipoyl-7-ADCA to 3-hydroxymethyl, to give the first product, 7-aminodeacetylcephalosporanic acid (7-ADAC); and3) an acetyltransferase gene whose expression product converts the 3-hydroxymethyl side chain to the 3-acetyloxymethyl side chain of 7-ACA.

Abstract: O-acylated cephalosporins are produced by reacting 3-hydroxymethylcephalosporins with an acyl donor containing at least three carbon atoms and an enzyme which is a Rodosporidium toruloides esterase, a wheat germ lipase, an Aspergillus niger lipase, an orange peel acetylesterase or a Bacillus subtilis esterase. A preferred enzyme is the esterase from Rodosporidium toruloides ATCC 10657. The enzyme can be used while in whole cells or in soluble or inmobilized form.

Abstract: Transformation of Cephalosporin C or its derivatives and salts into 7-aminocephalosporanic acid or its derivatives by an enzymatic two stage process with enzymes immobilized on a solid matrix.

Abstract: A process for the preparation of a compound of the formula ##STR1## wherein R.sub.1 is alkyl of 1 to 3 carbon atoms, R.sub.2 is alkyl of 1 to 12 carbon atoms, R.sub.3 is alkyl of 1 to 4 carbon atoms comprising reacting a compound of the formula ##STR2## wherein R.sub.4 is a remainder of an easily cleavable ether, K is a protected ketone function in the form of ketal, thioketal or mixed ketal and R.sub.1 has the above definition with a magnesium organic compound reagent of the formulaHalMg--CH.sub.2 --R.sub.2 Awherein Hal is a halogen and R.sub.2 is defined as above to obtain a compound of the formula ##STR3## wherein R.sub.1, R.sub.2 and K are defined as above, reacting the latter with an acylation agent to obtain a compound of the formula ##STR4## wherein R.sub.5 is alkyl of 1 to 6 carbon atoms or aryl of 6 to 10 carbon atoms and R.sub.1, R.sub.2 and K are defined as above, reacting the latter with an appropriate alkylation agent to obtain a compound of the formula ##STR5## wherein R.sub.1, R.sub.2, R.sub.

Abstract: A process for the production of an E. coli strain producing high yields of 7.beta.-(4-carboxybutanamido) cephalosporin acylase, comprises: (a) digesting the DNA of a microorganism whose DNA includes the sequence encoding 7.beta.-(4-carboxybutanamido) cephalosporin acylase, and forming a plasmidic library; (b) transforming, with the sequences in the plasmidic library, an auxotrophic E. coli host; (c) selecting for transformed E. coli hosts containing the acylase sequence by growth on a suitable medium; (d) isolating the vector containing the acylase sequence, digesting the vector, and ligating the DNA sequences obtained into an E. coli vector under control of an E. coli promoter; (e) repeating selection procedure of step (c) ; (f) using the vectors from the selected E. coli hosts to transform an E. coli host lacking substantial .beta.-lactamase activity. 7-Aminocephalosporanic acid and its derivatives can be prepared by reaction of substrates like 7.beta.-(4-carboxybutanamido) cephalosporanic acid with 7.beta.

Abstract: The present invention concerns a mutant cephalosporin C acylase derived from a precursor of the formula:A.sup.1-268 --X.sup.1 --Tyr--X.sup.2 --A.sup.272-304 --X.sup.3 --A.sup.306-773(SEQ ID NO:1), wherein:A.sup.1-268 is the same amino acid sequence as that from Thr.sup.1 to Gly.sup.268 of native CC acylase,A.sup.272-304 is the same amino acid sequence as that from Gln.sup.272 to Tyr.sup.304 of native CC acylase,A.sup.306-773 is the same amino acid sequence as that from Val.sup.306 to Ala.sup.773 of native CC acylase,X.sup.1 is Met or other amino acid,X.sup.2 is Ala or Tyr, andX.sup.3 is Cys or Ser,provided that when X.sup.1 is Met and X.sup.2 is Ala, X.sup.3 is Ser; and that the mutant cephalosporin C acylase has a property selected from the group consisting of higher enzymatic potency and higher processing efficiency, as compared to native CC acylase.

Abstract: 7-amino-cephalosporanic acids having the formula ##STR1## are obtained by submitting compounds having the following formula to enzymatic conversion: ##STR2## wherein R represents H, OH, O--CO--R", R" being an alkyl radical with from 1 to 4 carbon atoms, R.sup.1 represents a carboxylic group. The reaction is carried out in the presence of a microorganism selected from the Pseudomonas, Bacillus, Achromobacter genera, or in the presence of an enzyme, either free or immobilized, derived therefrom.

Abstract: An important intermediate for preparing cephalosporin antibiotics, 7-aminodesacetoxy cephalosporanic acid (7-ADCA), is prepared by a novel bioprocess in which a transformed Penicillium chrysogenum strain is cultured in the presence of an adipate feedstock to produce adipoyl-6-APA (6-amino penicillanic acid); and the in situ expression of an expandase gene, e.g., from Streptomyces clavuligerus, with which the P. chrysogenum has been transformed, converts the adipoly-6-APA by ring expansion to adipoyl-7-ADCA. The final product, 7-ADCA, is then prepared by cleavage of the adipoyl side chain using an adipoyl acylase. The entire synthesis, accordingly, is carried out using bioprocesses, and is efficient and economical.

Abstract: The enzymatic preparation of cephalosporanic derivatives, or their salts, ving the formula: ##STR1## wherein R is --CO--COOH or --COOH, R.sub.1 is H, OH, or --O--CO--R" and R" is an alkyl group with 1 to 4 carbon atoms, is carried out by the oxidative deamination of compounds, or their salts, having the formula: ##STR2## with an oxidase D-Aminoacid enzyme derived from Rhodotorula glutinis NCIMB 40412. The enzyme can be in a free or immobilized form.

Abstract: An improved technique and apparatus for automatic titration for determining the water content of a sample by a Karl Fischer (K.F.) reaction. Rather than requiring titration from one endpoint to another identical endpoint, each determination is made using a calibration relative to the water equivalent of a known amount of titrant together with the change in sensor output that occurs when a sample is added to the K.F. reagent. Improved accuracy and speed of analysis results.

Abstract: A cephalosporin C acylase from Pseudomonas diminuta N-176 is disclosed as well as the recombinant production of this enzyme in E. coli. The enzyme is characterized by the ability to catalyze the conversion of cephalosporin C, gultaryl 7-ACA, apidyl 7-ACA, succinyl 7-ACA, N-acetylcephalosporin C, N-benzoylcephalosporin C, and cephalothin into 7-aminocephalosporanic acid and is composed of an .alpha.-subunit with a molecular weight of 26,000 daltons and a .beta.-subunit with a molecular weight of 58,000 daltons.

Abstract: The present invention relates to a new method of protection of the carboxy group in the chemistry of the compounds of .beta.-lactam type. According to such a method, the carboxy group is protected by being transformed into its corresponding phenyl-acetoxy-methyl ester, which is then removed by an enzymatic route by means of penicillinacylase.In particular, if the .beta.-lactam compound also bears a phenyl-acetamidic substitutent, this latter is simultaneously hydrolysed during the same step of removal of the carboxy function protecting group.

Abstract: A process for the one-step conversion of cephalosporin C and derivatives thereof to the corresponding 7-aminocephalosporanic acid and derivatives comprising treating said cephalosporin C and derivatives with a cephalosporin C amidase derived from Arthrobacter viscosus ATCC 53594, or from any cephalosporin C amidase producing, or potentially producing descendants thereof, or from any expression of the genetic material of said Arthrobacter viscosus ATCC 53594, or any cephalosporin C amidase producing, or potentially producing descendants thereof.

Abstract: A process for preparing a 7-aminocephalosporanic acid compound which comprises treating a cephalosporin C compound with an enzyme-producing microorganism belonging to the genus Pseudomonas or a material obtained by subjecting the microorganism to chemical and/or physical treatment. A direct hydrolysis of the cephalosporin C compound into the 7-aminocephalosporanic acid compound and D-.alpha.-aminoadipic acid can be attained by the microorganism or material.

Abstract: Optically active cephalosporin analogs are produced by optically selective deacylation of an optically inactive acylated analog. The compounds are useful as intermediates in the preparation of optically active acylated antimicrobial agents.